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| - | {|{| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;"
| + | <html> |
| - | |- style="background:{{{color1|#ccccff}}};"
| + | <style type="text/css"> |
| - | |colspan="7"|{{{header|July}}}
| + | <!-- |
| - | |-
| + | h2 { |
| - | |- style="background:{{{color2|#eeeeff}}};"
| + | font-weight:bold; |
| - | |width="14%"|[[S|<span style="color:red;">S</span>]]
| + | } |
| - | |width="14%"|[[M]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[W]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[F]]
| + | |
| - | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]]
| + | |
| - | |-
| + | |
| - | |colspan ="4"|
| + | |
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]]
| + | |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]]
| + | |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|deepskyblue}}};"}}}>3</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|red}}};"}}}>4</span>]]
| + | |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]]
| + | |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]]
| + | |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]]
| + | |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]]
| + | |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]]
| + | |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|deepskyblue}}};"}}}>10</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|red}}};"}}}>11</span>]]
| + | |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]]
| + | |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]]
| + | |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]]
| + | |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]]
| + | |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]]
| + | |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|deepskyblue}}};"}}}>17</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|red}}};"}}}>18</span>]]
| + | |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]]
| + | |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]]
| + | |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>]]
| + | |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]]
| + | |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]]
| + | |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|deepskyblue}}};"}}}>24</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|red}}};"}}}>25</span>]]
| + | |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]]
| + | |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]]
| + | |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>]]
| + | |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]]
| + | |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]]
| + | |
| - | |[[{{{prefix|}}}31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}>31</span>]]
| + | |
| - | |-
| + | |
| - | |- style="background:{{{color3|#eeeeff}}};"
| + | |
| - | |colspan="7"|{{{footer|}}}
| + | |
| - | |}<noinclude>
| + | |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;"
| + | |
| - | |- style="background:{{{color1|#ccccff}}};"
| + | |
| - | |colspan="7"|{{{header|August}}}
| + | |
| - | |-
| + | |
| - | |- style="background:{{{color2|#eeeeff}}};"
| + | |
| - | |width="14%"|[[S|<span style="color:red;">S</span>]]
| + | |
| - | |width="14%"|[[M]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[W]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[F]]
| + | |
| - | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|red}}};"}}}>1</span>]]
| + | |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]]
| + | |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]]
| + | |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span>]]
| + | |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]]
| + | |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]]
| + | |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|deepskyblue}}};"}}}>7</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|red}}};"}}}>8</span>]]
| + | |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]]
| + | |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]]
| + | |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span>]]
| + | |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]]
| + | |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]]
| + | |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|deepskyblue}}};"}}}>14</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span>]]
| + | |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]]
| + | |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]]
| + | |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span>]]
| + | |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]]
| + | |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]]
| + | |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|deepskyblue}}};"}}}>21</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|red}}};"}}}>22</span>]]
| + | |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]]
| + | |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span>]]
| + | |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span>]]
| + | |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]]
| + | |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]]
| + | |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|deepskyblue}}};"}}}>28</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|red}}};"}}}>29</span>]]
| + | |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]]
| + | |
| - | |[[{{{prefix|}}}31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|inherit}}};"}}}>31</span>]]
| + | |
| - | |colspan="4"|
| + | |
| - | |- style="background:{{{color3|#eeeeff}}};"
| + | |
| - | |colspan="7"|{{{footer|}}}
| + | |
| - | |}<noinclude>
| + | |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;"
| + | |
| - | |- style="background:{{{color1|#ccccff}}};"
| + | |
| - | |colspan="7"|{{{header|September}}}
| + | |
| - | |-
| + | |
| - | |- style="background:{{{color2|#eeeeff}}};"
| + | |
| - | |width="14%"|[[S|<span style="color:red;">S</span>]]
| + | |
| - | |width="14%"|[[M]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[W]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[F]]
| + | |
| - | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]]
| + | |
| - | |-
| + | |
| - | |colspan ="3"|
| + | |
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]]
| + | |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]]
| + | |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]]
| + | |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|deepskyblue}}};"}}}>4</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|red}}};"}}}>5</span>]]
| + | |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]]
| + | |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]]
| + | |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]]
| + | |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]]
| + | |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]]
| + | |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}>11</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|red}}};"}}}>12</span>]]
| + | |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]]
| + | |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]]
| + | |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]]
| + | |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]]
| + | |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]]
| + | |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}>18</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|red}}};"}}}>19</span>]]
| + | |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]]
| + | |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>]]
| + | |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]]
| + | |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]]
| + | |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span>]]
| + | |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|deepskyblue}}};"}}}>25</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|red}}};"}}}>26</span>]]
| + | |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]]
| + | |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>]]
| + | |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]]
| + | |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]]
| + | |
| - | |colspan="2"|
| + | |
| - | |- style="background:{{{color3|#eeeeff}}};"
| + | |
| - | |colspan="7"|{{{footer|}}}
| + | |
| - | |}<noinclude>
| + | |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;"
| + | |
| - | |- style="background:{{{color1|#ccccff}}};"
| + | |
| - | |colspan="7"|{{{header|October}}}
| + | |
| - | |-
| + | |
| - | |- style="background:{{{color2|#eeeeff}}};"
| + | |
| - | |width="14%"|[[S|<span style="color:red;">S</span>]]
| + | |
| - | |width="14%"|[[M]]
| + | |
| - | |width="14%"|[[T]]
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| - | |width="14%"|[[W]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[F]]
| + | |
| - | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]]
| + | |
| - | |-
| + | |
| - | |colspan ="5"|
| + | |
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]]
| + | |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|deepskyblue}}};"}}}>2</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|red}}};"}}}>3</span>]]
| + | |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span>]]
| + | |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]]
| + | |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]]
| + | |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]]
| + | |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]]
| + | |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|deepskyblue}}};"}}}>9</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|red}}};"}}}>10</span>]]
| + | |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span>]]
| + | |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]]
| + | |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]]
| + | |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]]
| + | |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]]
| + | |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|deepskyblue}}};"}}}>16</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|red}}};"}}}>17</span>]]
| + | |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span>]]
| + | |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]]
| + | |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]]
| + | |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>]]
| + | |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]]
| + | |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|deepskyblue}}};"}}}>23</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|red}}};"}}}>24</span>]]
| + | |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span>]]
| + | |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]]
| + | |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]]
| + | |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>]]
| + | |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]]
| + | |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|deepskyblue}}};"}}}>30</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|red}}};"}}}>31</span>]]
| + | |
| - | |colspan="6"|
| + | |
| - | |- style="background:{{{color3|#eeeeff}}};"
| + | |
| - | |colspan="7"|{{{footer|}}}
| + | |
| - | |}<noinclude>
| + | |
| - | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|150px}}}; margin-left:1em; text-align:center;"
| + | |
| - | |- style="background:{{{color1|#ccccff}}};"
| + | |
| - | |colspan="7"|{{{header|November}}}
| + | |
| - | |-
| + | |
| - | |- style="background:{{{color2|#eeeeff}}};"
| + | |
| - | |width="14%"|[[S|<span style="color:red;">S</span>]]
| + | |
| - | |width="14%"|[[M]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[W]]
| + | |
| - | |width="14%"|[[T]]
| + | |
| - | |width="14%"|[[F]]
| + | |
| - | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]]
| + | |
| - | |-
| + | |
| - | |colspan ="1"|
| + | |
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]]
| + | |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]]
| + | |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]]
| + | |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span>]]
| + | |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]]
| + | |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|deepskyblue}}};"}}}>6</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|red}}};"}}}>7</span>]]
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| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]]
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| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]]
| + | |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]]
| + | |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span>]]
| + | |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]]
| + | |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|deepskyblue}}};"}}}>13</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|red}}};"}}}>14</span>]]
| + | |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]]
| + | |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]]
| + | |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]]
| + | |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span>]]
| + | |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]]
| + | |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|deepskyblue}}};"}}}>20</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|red}}};"}}}>21</span>]]
| + | |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]]
| + | |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]]
| + | |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span>]]
| + | |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span>]]
| + | |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]]
| + | |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|deepskyblue}}};"}}}>27</span>]]
| + | |
| - | |-
| + | |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|red}}};"}}}>28</span>]]
| + | |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]]
| + | |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]]
| + | |
| - | |colspan="4"|
| + | |
| - | |- style="background:{{{color3|#eeeeff}}};"
| + | |
| - | |colspan="7"|{{{footer|}}}
| + | |
| - | |}<noinclude>
| + | |
| | | | |
| | + | #contentSub { |
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| | + | } |
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| | + | #search-controls { |
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| | + | #top-section { |
| | + | height:25px; |
| | + | width:1000px; |
| | + | color: blue; |
| | + | background-color: transparent; |
| | + | background-image: url(https://static.igem.org/mediawiki/2010/f/f1/Osaka_top_section.jpg) |
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| | + | #p-logo { |
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| | + | #menubar { |
| | + | top:0px; |
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| | + | background-color: transparent; |
| | + | background-image: url(https://static.igem.org/mediawiki/2010/a/af/Osaka_sand2.jpg); |
| | + | padding-bottom: 20px; |
| | + | padding-left:0px; |
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| | | | |
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| | + | position: relative; |
| | + | width:1000px; |
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| | + | width:1000px; |
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| | + | margin:0; |
| | + | background-color:#0342ff; |
| | + | |
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| | + | .banner{ |
| | + | width:840px; |
| | + | float:left; |
| | + | padding:0; |
| | + | margin:0; |
| | + | background-color:black; |
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| | + | .bottom{ |
| | + | width:1000px;} |
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| | + | width:840px; |
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| | + | .contents2{ |
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| | + | width:750px; |
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| | + | padding:0 10px; |
| | + | margin:0; |
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| | + | } |
| | | | |
| | + | .menu{ |
| | + | width:160px; |
| | + | float:left; |
| | + | background-color:white; |
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| | + | } |
| | + | .clear{ |
| | + | clear:both;} |
| | + | .clear hr{ |
| | + | display:none;} |
| | + | .bold { |
| | + | font-weight:bold; |
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| | + | </style> |
| | + | </head> |
| | | | |
| | | | |
| | + | <body> |
| | | | |
| | + | <div class="wrap"> |
| | + | <div class="top"> |
| | + | <div class="banner"> |
| | + | <img src="https://static.igem.org/mediawiki/2010/d/d6/Osaka_banner3.jpg" width="1000" height="250" alt="iGEM Osaka banner" usemap="#banner_map"> |
| | | | |
| | + | <map name="banner_map"> |
| | + | <area shape="rect" coords="540,180,750,220" href="https://2010.igem.org" alt="iGEM 2010 home"/> |
| | + | <area shape="rect" coords="430,230,750,270" href="http://www.osaka-u.ac.jp/en" alt="Osaka University"/> |
| | + | <area shape="circle" coords="100,200,85" href="https://2010.igem.org/Team:Osaka" alt="iGEM Osaka home"/> |
| | + | </map> |
| | | | |
| - | ---- | + | </div><!-- banner --> |
| | + | <div class="clear"><hr></div> |
| | + | </div><!-- top --> |
| | | | |
| | + | <div class="bottom"> |
| | + | <div class="menu"> |
| | + | </html> |
| | + | {{Osaka_menu}} |
| | + | <html> |
| | + | </div> |
| | | | |
| - | ==Notebook==
| |
| - | ===July 29 (Thu)===
| |
| - | Attendance: Torigata, Takino, Teoh, Yasumoto, Kakuda, Saka, Tamura
| |
| - | # Safety lecture for junior members.
| |
| - | # Preparation of LB agar plates (26 Amp, 25 Kan, 25 Cam).
| |
| | | | |
| - | ===July 31 (Sat)===
| |
| - | Attendance: Miyatake, Hirayama, Torigata, Teoh, Tadashi, Yasumoto, Kakuda, Saka
| |
| - | # Meeting
| |
| - | #* Summer project schedule
| |
| - | #* List of genes to clone
| |
| | | | |
| - | ===August 2 (Mon)===
| |
| - | Attendance: Torigata, Takino, Teoh, Tadashi, Yasumoto, Saka
| |
| - | # Culture medium preparation
| |
| - | #* LB agar plates (49 antibiotic-less plates)
| |
| - | #* LB liquid medium (500 ml)
| |
| - | # Competent cells preparation - Nojima Method
| |
| - | #* SOB medium (MgCl2 not yet added) -> stored at 4˚C
| |
| - | #* TB buffer -> stored at 4˚C
| |
| - | #* Single-colony streaking of E. coli (strain DH5α) on 2 LB agar plates -> 37˚C incubation o/n
| |
| - | Tomorrow we shall prepare the remainder of the reagents and inoculate SOB with o/n-incubated E. coli.
| |
| | | | |
| - | ===August 3 (Tue)=== | + | <html> |
| - | Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
| + | <div class="right"> |
| - | # Competent cells preparation (continued)
| + | <div class="contents"> |
| - | #* Preparation of glucose solution for making SOC medium.
| + | <div style="margin:40px 0"> |
| - | #* Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
| + | </html> |
| - | #* (Night) Transfer from pre-culture to growth culture.
| + | ==Notebook== |
| - | | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| - | ===August 4 (Wed)=== | + | |- style="background:{{{color1|#ccccff}}};" |
| - | Attendance: Nakamura, Saka, Kakuda, Yasumoto, Torigata, Teoh
| + | |colspan="8"|{{{header|July / August}}} |
| - | # OD measurements throughout the day till required OD (0.3~0.7) was obtained.
| + | |
| - | # Completion of competent cells according to protocol.
| + | |
| - | | + | |
| - | ===August 5 (Thu)=== | + | |
| - | Attendance: Nakamura, Yasumoto, Saka, Kakuda, Takino
| + | |
| - | # Transformation of Registry parts: | + | |
| - | {| | + | |
| - | !ID!!Part Name!!Resistance!!Description
| + | |
| | |- | | |- |
| - | |2-20J||<bbpart>BBa_K118023</bbpart>||A||''C. fermi'' endocellulase Cen A coding | + | |- style="background:{{{color2|#eeeeff}}};" |
| | + | |width=100px|Week |
| | + | |width="14%"|<span style="color:red;">S</span> |
| | + | |width="14%"|M |
| | + | |width="14%"|T |
| | + | |width="14%"|W |
| | + | |width="14%"|T |
| | + | |width="14%"|F |
| | + | |width="14%"|<span style="color:deepskyblue;">S</span> |
| | |- | | |- |
| - | |2-20H||<bbpart>BBa_K118022</bbpart>||A||''C. fermi'' exocellulase Cex coding | + | |<span style>[[Team:Osaka/week1|week1]]</span> |
| | + | |<span style={{{s25|"color:{{{c25|red}}};"}}}>25</span> |
| | + | |<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span> |
| | + | |<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span> |
| | + | |<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span> |
| | + | |[[Team:Osaka/week1#July 29 (Thu)|29]]<span style={{{s29|"color:{{{c29|inherit}}};"}}}></span> |
| | + | |<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span> |
| | + | |[[Team:Osaka/week1#July 31 (Sat)|31]]<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |1-2M||<bbpart>BBa_B0034</bbpart>||A||RBS | + | |<span style>[[Team:Osaka/week2|week2]]</span> |
| | + | |<span style={{{s1|"color:{{{c1|red}}};"}}}>1</span> |
| | + | |[[Team:Osaka/week2#August 2 (Mon)|2]]<span style={{{s2|"color:{{{c2|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week2#August 3 (Tue)|3]]<span style={{{s3|"color:{{{c3|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week2#August 4 (Wed)|4]]<span style={{{s4|"color:{{{c4|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week2#August 5 (Thu)|5]]<span style={{{s5|"color:{{{c5|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week2#August 6 (Fri)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week2#August 7 (Sat)|7]]<span style={{{s7|"color:{{{c7|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |1-13D||<bbpart>BBa_B0010</bbpart>||A||terminator | + | |<span style>[[Team:Osaka/week3|week3]]</span> |
| | + | |[[Team:Osaka/week3#August 8 (Sun)|8]]<span style={{{s8|"color:{{{c8|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week3#August 9 (Mon)|9]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week3#August 10 (Tue)|10]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week3#August 11 (Wed)|11]]<span style={{{s11|"color:{{{c11|inherit}}};"}}}></span> |
| | + | |<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span> |
| | + | |<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span> |
| | + | |<span style={{{s14|"color:{{{c14|deepskyblue}}};"}}}>14</span> |
| | |- | | |- |
| - | |1-1D||<bbpart>BBa_R0010</bbpart>||A||promoter | + | |<span style>[[Team:Osaka/week4|week4]]</span> |
| | + | |<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span> |
| | + | |[[Team:Osaka/week4#August 16 (Mon)|16]]<span style={{{s16|"color:{{{c16|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week4#August 17 (Tue)|17]]<span style={{{s17|"color:{{{c17|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week4#August 18 (Wed)|18]]<span style={{{s18|"color:{{{c18|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week4#August 19 (Thu)|19]]<span style={{{s19|"color:{{{c19|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week4#August 20 (Fri)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week4#August 21 (Sat)|21]]<span style={{{s21|"color:{{{c21|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |1-18F||<bbpart>BBa_E1010</bbpart>||K||RFP coding
| + | |<span style>[[Team:Osaka/week5|week5]]</span> |
| - | |}
| + | |[[Team:Osaka/week5#August 22 (Sun)|22]]<span style={{{s22|"color:{{{c22|red}}};"}}}></span> |
| - | Note: 'ID' is an internal identifier used by wet work members to simplify labeling etc. In the case of iGEM distribution parts it usually refers to the plate location.
| + | |[[Team:Osaka/week5#August 23 (Mon)|23]]<span style={{{s23|"color:{{{c23|inherit}}};"}}}></span> |
| - | | + | |[[Team:Osaka/week5#August 24 (Tue)|24]]<span style={{{s24|"color:{{{c24|inherit}}};"}}}></span> |
| - | ===August 6 (Fri)===
| + | |[[Team:Osaka/week5#August 25 (Wed)|25]]<span style={{{s25|"color:{{{c25|inherit}}};"}}}></span> |
| - | Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
| + | |[[Team:Osaka/week5#August 26 (Thu)|26]]<span style={{{s26|"color:{{{c26|inherit}}};"}}}></span> |
| - | # Colony check
| + | |[[Team:Osaka/week5#August 27 (Fri)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span> |
| - | #* All transformed cells produced colonies! | + | |[[Team:Osaka/week5#August 28 (Sat)|28]]<span style={{{s28|"color:{{{c28|deepskyblue}}};"}}}></span> |
| - | #* Non-transformed cells (negative controls) did not grow on Amp, Kan or Cam plates -> confirmed lack of natural antibiotic resistance
| + | |
| - | # Colonies transferred to 3ml LB growth medium & incubated o/n at 37˚C | + | |
| - | | + | |
| - | ===August 7 (Sat)===
| + | |
| - | Attendance: Nakamura, Saka, Yasumoto, Takino
| + | |
| - | # Miniprep of o/n-incubated colonies (2-20J, 2-20H, 1-2M, 1-13D, 1-1D, 1-18F) using Sigma-Aldrich Plasmid Miniprep Kit
| + | |
| - | # Transformation of construction plasmids | + | |
| - | {| | + | |
| - | !ID!!Part Name!!Resistance!!Description
| + | |
| | |- | | |- |
| - | |1-1C||<bbpart>pSB1A3</bbpart>||A||construction plasmid containing mRFP coding device (<bbpart>BBa_J04450</bbpart>) | + | |<span style>[[Team:Osaka/week6|week6]]</span> |
| | + | |[[Team:Osaka/week6#August 29 (Sun)|29]]<span style={{{s29|"color:{{{c29|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week6#August 30 (Mon)|30]]<span style={{{s30|"color:{{{c30|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week6#August 31 (Tue)|31]]<span style={{{s31|"color:{{{c31|inherit}}};"}}}></span> |
| | + | |colspan="4"| |
| | + | |- style="background:{{{color3|#eeeeff}}};" |
| | + | |colspan="7"|{{{footer|}}} |
| | + | |}<noinclude> |
| | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| | + | |- style="background:{{{color1|#ccccff}}};" |
| | + | |colspan="8"|{{{header|September}}} |
| | |- | | |- |
| - | |1-3A||<bbpart>pSB1C3</bbpart>||C||<nowiki>(" ")</nowiki> | + | |- style="background:{{{color2|#eeeeff}}};" |
| | + | |width=100px|Week |
| | + | |width="14%"|<span style="color:red;">S</span> |
| | + | |width="14%"|M |
| | + | |width="14%"|T |
| | + | |width="14%"|W |
| | + | |width="14%"|T |
| | + | |width="14%"|F |
| | + | |width="14%"|<span style="color:deepskyblue;">S</span> |
| | |- | | |- |
| - | |1-5A||<bbpart>pSB1K3</bbpart>||K||<nowiki>(" ")</nowiki>
| + | |<span style>[[Team:Osaka/week6|week6]]</span> |
| - | |} | + | |<span style></span> |
| - | # Meeting
| + | |<span style></span> |
| - | | + | |<span style></span> |
| - | ===August 8 (Sun)===
| + | |[[Team:Osaka/week6#September 1 (Wed)|1]]<span style={{{s1|"color:{{{c1|inherit}}};"}}}></span> |
| - | Attendance: Nakamura, Yasumoto
| + | |[[Team:Osaka/week6#September 2 (Thu)|2]]<span style={{{s2|"color:{{{c2|inherit}}};"}}}></span> |
| - | # Colony check
| + | |[[Team:Osaka/week6#September 3 (Fri)|3]]<span style={{{s3|"color:{{{c3|inherit}}};"}}}></span> |
| - | #*All parts successfully transformed
| + | |[[Team:Osaka/week6#September 4 (Sat)|4]]<span style={{{s4|"color:{{{c4|deepskyblue}}};"}}}></span> |
| - | # Transfer to LB culture medium
| + | |
| - | | + | |
| - | ===August 9 (Mon)===
| + | |
| - | # Miniprep of 1-1C
| + | |
| - | # Restriction digests of 2-20H, 2-20J, 1-1D, 1-18F, 1-1C, 1-13D, 1-2M
| + | |
| - | #* (WHICH ENZYMES?)
| + | |
| - | # Gel electrophoresis of digests ("cut check")
| + | |
| - | #* 2-20H, 2-20J, 1-1C -> OK
| + | |
| - | #* 1-1D, 1-18F, 1-13D, 1-2M -> not cut (single band, MW approx. equal to vector + insert)
| + | |
| - | # Transfer of 1-3A, 1-5A colonies to solution culture (repeat)
| + | |
| - | #*''Yesterday's inoculated culture mediums contained the wrong antibiotics!''
| + | |
| - | # Transformation of <bbpart>BBa_I13521</bbpart>, <bbpart>BBa_I13522</bbpart>, <bbpart>BBa_I13600</bbpart>, <bbpart>BBa_K204031</bbpart>, <bbpart>BBa_K204051</bbpart>, <bbpart>BBa_K204032</bbpart> from last year's stock
| + | |
| - | | + | |
| - | ===August 10 (Tue)===
| + | |
| - | # Miniprep of 1-3A, 1-5A
| + | |
| - | # Restriction digests of 1-3A, 1-5A
| + | |
| - | # Gel electrophoresis
| + | |
| - | ## 1st run: 1-3A, 1-5A (newly miniprepped), 1-1D, 1-18F, 1-13D, 1-2M (repeat)
| + | |
| - | ##* 1-3A, 1-5A -> OK; others -> not cut (''again'')
| + | |
| - | ## 2nd run: 1-1D, 1-18F, 1-13D, 1-2M (2nd repeat)
| + | |
| - | ##* all parts not cut | + | |
| - | | + | |
| - | ===August 11 (Wed)===
| + | |
| - | # Miniprep of last year's parts transformed on Monday
| + | |
| - | # Gel electrophoresis of 1-1D, 1-18F, 1-13D, 1-2M (3rd repeat!!!)
| + | |
| - | #* all 4 not cut... AGAIN
| + | |
| - | #* so far all restriction digests involving XbaI seem to have failed; problem with enzyme stock?
| + | |
| - | #* will try with different set of restriction enzymes next week
| + | |
| - | # Sent miniprepped last year's parts to ECUST team in Shanghai, China
| + | |
| - | | + | |
| - | ===August 16 (Mon)===
| + | |
| - | # Restriction digests
| + | |
| - | #* 1-13D, 1-2M, 1-1D, 1-18F with EcoRI, PstI to check for point mutation(s) affecting XbaI site
| + | |
| - | #* 2-20J with XbaI, PstI to check/confirm XbaI activity
| + | |
| - | # Gel electrophoresis of digests
| + | |
| - | #* (RESULTS?)
| + | |
| - | # Colony pick-up & transfer to solution culture (repeat; 2 each): 1-13D, 1-2M, 1-1D, 1-18F
| + | |
| - | #* 3ml LB liquid medium; 3μl Amp or 0.6μl Kan solution added | + | |
| - | | + | |
| - | ===August 17 (Tue)===
| + | |
| - | # Mini-prep of 1-13D, 1-2M, 1-1D, 1-18F parts inoculated yesterday (2 of each)
| + | |
| - | # 'Cut check' (restriction digest + gel electrophoresis to confirm insert) of the miniprepped parts:
| + | |
| - | #* 1-1D, 1-18F with EcoRI, SpeI
| + | |
| - | #* 1-13D, 1-2M with XbaI, PstI
| + | |
| - | #* (RESULTS?)
| + | |
| - | # Transformation of secretion tag parts using 25μl of competent cells each
| + | |
| - | {| | + | |
| - | !ID!!Part Name!!Resistance!!Description
| + | |
| | |- | | |- |
| - | |2-22P||<bbpart>BBa_K103006</bbpart>||A||OmpA outer membrane protein + linker | + | |<span style>[[Team:Osaka/week7|week7]]</span> |
| | + | |[[Team:Osaka/week7#September 5 (Sun)|5]]<span style={{{s5|"color:{{{c5|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 6 (Mon)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 7 (Tue)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 8 (Wed)|8]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 9 (Thu)|9]]<span style={{{s9|"color:{{{c9|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 10 (Fri)|10]]<span style={{{s10|"color:{{{c10|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week7#September 11 (Sat)|11]]<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |1-2J||<bbpart>BBa_J32015</bbpart>||A,K||PelB leader sequence
| + | |<span style>[[Team:Osaka/week8|week8]]</span> |
| - | |} | + | |[[Team:Osaka/week8#September 12 (Sun)|12]]<span style={{{s12|"color:{{{c12|red}}};"}}}></span> |
| - | | + | |[[Team:Osaka/week8#September 13 (Mon)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span> |
| - | ===August 18 (Wed)===
| + | |[[Team:Osaka/week8#September 14 (Tue)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></span> |
| - | iGEM Japan Meet-Up in Kyoto
| + | |[[Team:Osaka/week8#September 15 (Wed)|15]]<span style={{{s15|"color:{{{c15|inherit}}};"}}}></span> |
| - | Attendance: Nakamura, Yasumoto, Saka, Kakuda
| + | |[[Team:Osaka/week8#September 16 (Thu)|16]]<span style={{{s16|"color:{{{c16|inherit}}};"}}}></span> |
| - | | + | |[[Team:Osaka/week8#September 17 (Fri)|17]]<span style={{{s17|"color:{{{c17|inherit}}};"}}}></span> |
| - | ===August 19 (Thu)===
| + | |[[Team:Osaka/week8#September 18 (Sat)|18]]<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}></span> |
| - | # Transfer of 2-22P, 1-2J to solution culture | + | |
| - | # Gel electrophoresis of digests from 'cut check' products from Tuesday
| + | |
| - | #* repeat run, but each digest together with undigested plasmid DNA)
| + | |
| - | #* 2% agarose gel instead of the usual 1%
| + | |
| - | #* (RESULTS?)
| + | |
| - | # Gel electrophoresis of 1-1D digest only
| + | |
| - | #* (RESULT?)
| + | |
| - | # Multiple restriction digests of 1-1D to check for problems at restriction sites
| + | |
| - | #* tried the following: EcoRI only; SpeI only; EcoRI + SpeI
| + | |
| - | # Night: miniprep of 2-22P, 1-2J inoculated in the morning
| + | |
| - | | + | |
| - | ===August 20 (Fri)===
| + | |
| - | # Gel electrophoresis of 1-1D and its digests
| + | |
| - | #* (RESULTS?) | + | |
| - | # 'Cut check' of parts miniprepped the night before
| + | |
| - | #* both 2-22P & 1-2J cut with XbaI, PstI
| + | |
| - | #* enzyme inactivation at 80℃, 20min
| + | |
| - | #* (RESULTS?)
| + | |
| - | # Restriction digest of 2-20J (WHICH ENZYMES?)
| + | |
| - | # Ligation according to 3A assembly method: 2-20J or 2-20H + 2-22P or 1-2J; 1-3A as vector
| + | |
| - | #* reaction mixture: 2μl of each plasmid, 2μl ligase buffer, 1μl T4 DNA ligase, water to make 20μl total
| + | |
| - | #* reaction at room temperature for 10min; ligase inactivation at 80℃ for 20min | + | |
| - | # Transformation of ligated parts using 50μl of competent cells each; 2μl ligation product; 150μl Chloramphenicol spread on agar plates before inoculation with pre-incubation mix
| + | |
| - | | + | |
| - | ===August 21 (Sat)===
| + | |
| - | # Transfer of white (non-RFP) colonies from yesterday's transformation to 3ml LB liquid medium added with 1μl Cam solution | + | |
| - | #* ''we later realized that the upstream and downstream parts had been mixed up in these ligations, so the ligation products (and these corresponding solution cultures) were discarded''
| + | |
| - | # 3A assembly ligation: 1-1D as upstream, 1-2M as downstream, 1-3A as vector
| + | |
| - | #* ligation product designated as <1>; Chloramphenicol resistance
| + | |
| - | #* same ligation mix composition as yesterday's
| + | |
| - | # Transformation of <1> with pre-incubation for 1.5hr instead of 1hr
| + | |
| - | | + | |
| - | ===August 22 (Sun)===
| + | |
| - | # Transfer of <1> to culture solution; incubation at 30℃ (''why??'')
| + | |
| - | # Transformation of the following parts: | + | |
| - | {| | + | |
| - | !ID!!Part Name!!Resistance!!Description
| + | |
| | |- | | |- |
| - | |2-4A||<bbpart>BBa_J63005</bbpart>||A||yeast ADH1 promoter | + | |<span style>[[Team:Osaka/week9|week9]]</span> |
| | + | |[[Team:Osaka/week9#September 19 (Sun)|19]]<span style={{{s19|"color:{{{c19|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 20 (Mon)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 21 (Tue)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 22 (Wed)|22]]<span style={{{s22|"color:{{{c22|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 23 (Thu)|23]]<span style={{{s23|"color:{{{c23|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 24 (Fri)|24]]<span style={{{s24|"color:{{{c24|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week9#September 25 (Sat)|25]]<span style={{{s25|"color:{{{c25|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |F1||N/A||A||beta-galactosidase from Edinburgh team | + | |<span style>[[Team:Osaka/week10|week10]]</span> |
| | + | |[[Team:Osaka/week10#September 26 (Sun)|26]]<span style={{{s26|"color:{{{c26|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week10#September 27 (Mon)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week10#September 28 (Tue)|28]]<span style={{{s28|"color:{{{c28|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week10#September 29 (Wed)|29]]<span style={{{s29|"color:{{{c29|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week10#September 30 (Thu)|30]]<span style={{{s30|"color:{{{c30|inherit}}};"}}}></span> |
| | + | |colspan="2"| |
| | + | |- style="background:{{{color3|#eeeeff}}};" |
| | + | |colspan="7"|{{{footer|}}} |
| | + | |}<noinclude> |
| | + | {| class="toccolours" style="float:{{{float|left}}}; width:{{{width|240px}}}; text-align:center;" |
| | + | |- style="background:{{{color1|#ccccff}}};" |
| | + | |colspan="8"|{{{header|October}}} |
| | |- | | |- |
| - | |F2||N/A||C||RBS + F1 | + | |- style="background:{{{color2|#eeeeff}}};" |
| | + | |width=100px|Week |
| | + | |width="14%"|<span style="color:red;">S</span> |
| | + | |width="14%"|M |
| | + | |width="14%"|T |
| | + | |width="14%"|W |
| | + | |width="14%"|T |
| | + | |width="14%"|F |
| | + | |width="14%"|<span style="color:deepskyblue;">S</span> |
| | |- | | |- |
| - | |F3||N/A||C||Lac promoter + RBS + F1 | + | |<span style>[[Team:Osaka/week10|week10]]</span> |
| - | |} | + | |<span style></span> |
| - | #* O/N incubation at 37℃ as per normal
| + | |<span style></span> |
| - | | + | |<span style></span> |
| - | ===August 23 (Mon)===
| + | |<span style></span> |
| - | # Transfer of 2-4A, F1, F2, F3 transformed yesterday to solution culture
| + | |<span style></span> |
| - | # Miniprep & 'Cut check' of <1>
| + | |[[Team:Osaka/week10#October 1 (Fri)|1]]<span style={{{s1|"color:{{{c1|inherit}}};"}}}></span> |
| - | #* cut at EcoRI, SpeI | + | |[[Team:Osaka/week10#October 2 (Sat)|2]]<span style={{{s2|"color:{{{c2|deepskyblue}}};"}}}></span> |
| - | #* gel run with DNA ladder, digested plasmid, undigested plasmid
| + | |
| - | #* (RESULTS?)
| + | |
| - | # Transfer of 3 more colonies of <1> to liquid solution (''to store as glycerol stock - see Tue notes'')
| + | |
| - | # Transformation of the following registry parts
| + | |
| - | {| | + | |
| - | !ID!!Part Name!!Resistance!!Description
| + | |
| | |- | | |- |
| - | |2-2O||<bbpart>J63003</bbpart>||A||yeast Kozak sequence | + | |<span style>[[Team:Osaka/week11|week11]]</span> |
| | + | |[[Team:Osaka/week11#October 3 (Sun)|3]]<span style={{{s3|"color:{{{c3|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 4 (Mon)|4]]<span style={{{s4|"color:{{{c4|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 5 (Tue)|5]]<span style={{{s5|"color:{{{c5|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 6 (Wed)|6]]<span style={{{s6|"color:{{{c6|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 7 (Thu)|7]]<span style={{{s7|"color:{{{c7|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 8 (Fri)|8]]<span style={{{s8|"color:{{{c8|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week11#October 9 (Sat)|9]]<span style={{{s9|"color:{{{c9|deepskyblue}}};"}}}></span> |
| | |- | | |- |
| - | |3-11I||<bbpart>K105027</bbpart>||A||'cyc100' minimal promoter | + | |<span style>[[Team:Osaka/week12|week12]]</span> |
| - | |} | + | |[[Team:Osaka/week12#October 10 (Sun)|10]]<span style={{{s10|"color:{{{c10|red}}};"}}}></span> |
| - | | + | |[[Team:Osaka/week12#October 11 (Mon)|11]]<span style={{{s11|"color:{{{c11|inherit}}};"}}}></span> |
| - | ===August 24 (Tue)=== | + | |[[Team:Osaka/week12#October 12 (Tue)|12]]<span style={{{s12|"color:{{{c12|inherit}}};"}}}></span> |
| - | # Colony check of part transformed yesterday: both 2-2O & 3-11I produced >100 colonies | + | |[[Team:Osaka/week12#October 13 (Wed)|13]]<span style={{{s13|"color:{{{c13|inherit}}};"}}}></span> |
| - | #* transfer to solution culture | + | |[[Team:Osaka/week12#October 14 (Thu)|14]]<span style={{{s14|"color:{{{c14|inherit}}};"}}}></span> |
| - | # Miniprep of 2-4A, F1, F2, F3 followed by 'cut check' with EcoRI, SpeI | + | |[[Team:Osaka/week12#October 15 (Fri)|15]]<span style={{{s15|"color:{{{c15|inherit}}};"}}}></span> |
| - | #* (RESULTS) | + | |[[Team:Osaka/week12#October 16 (Sat)|16]]<span style={{{s16|"color:{{{c16|deepskyblue}}};"}}}></span> |
| - | # Transfer of F1 to solution culture (''why?'') | + | |- |
| - | # Preparation of glycerol stock of cell culture containing <1> (''why?'') | + | |<span style>[[Team:Osaka/week13|week13]]</span> |
| - | #* 200ml of culture solution mixed with 100ml of 50% glycerol | + | |[[Team:Osaka/week13#October 17 (Sun)|17]]<span style={{{s17|"color:{{{c17|red}}};"}}}></span> |
| - | #* stored at -80℃
| + | |[[Team:Osaka/week13#October 18 (Mon)|18]]<span style={{{s18|"color:{{{c18|inherit}}};"}}}></span> |
| - | | + | |[[Team:Osaka/week13#October 19 (Tue)|19]]<span style={{{s19|"color:{{{c19|inherit}}};"}}}></span> |
| - | ===August 25 (Wed)=== | + | |[[Team:Osaka/week13#October 20 (WEd)|20]]<span style={{{s20|"color:{{{c20|inherit}}};"}}}></span> |
| - | # Miniprep of parts in solution culture: 2-2O, 3-11I, F1
| + | |[[Team:Osaka/week13#October 21 (Thu)|21]]<span style={{{s21|"color:{{{c21|inherit}}};"}}}></span> |
| - | # Cut check of 3-11I & F1 with EcoRI, SpeI
| + | |[[Team:Osaka/week13#October 22 (Fri)|22]]<span style={{{s22|"color:{{{c22|inherit}}};"}}}></span> |
| - | #* (RESULTS?) | + | |[[Team:Osaka/week13#October 23 (Sat)|23]]<span style={{{s23|"color:{{{c23|deepskyblue}}};"}}}></span> |
| | + | |- |
| | + | |<span style>[[Team:Osaka/week14|week14]]</span> |
| | + | |[[Team:Osaka/week14#October 24 (Sun)|24]]<span style={{{s24|"color:{{{c24|red}}};"}}}></span> |
| | + | |[[Team:Osaka/week14#October 25 (Mon)|25]]<span style={{{s25|"color:{{{c25|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week14#October 26 (Tue)|26]]<span style={{{s26|"color:{{{c26|inherit}}};"}}}></span> |
| | + | |[[Team:Osaka/week14#October 27 (Wed)|27]]<span style={{{s27|"color:{{{c27|inherit}}};"}}}></span> |
| | + | |<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span> |
| | + | |<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span> |
| | + | |<span style={{{s30|"color:{{{c30|deepskyblue}}};"}}}>30</span> |
| | + | |- |
| | + | |<span style>[[Team:Osaka/week15|week15]]</span> |
| | + | |[[{{{prefix|}}}10.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|red}}};"}}}>31</span>]] |
| | + | |colspan="6"| |
| | + | |- style="background:{{{color3|#eeeeff}}};" |
| | + | |colspan="7"|{{{footer|}}} |
| | + | |}<noinclude> |
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| | </div> | | </div> |
| | + | </div> |
| | + | </div> |
| | + | <div class="clear"><hr></div> |
| | + | </html> |
| | + | {{Osaka_foot}} |
| | + | <html> |
| | + | |
| | </html> | | </html> |
"
