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Team:Osaka/Notebook
From 2010.igem.org
(Difference between revisions)
| Line 13: | Line 13: | ||
|- | |- | ||
|colspan ="4"| | |colspan ="4"| | ||
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]] | + | |[[{{{prefix|}}}7.1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]] |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] | + | |[[{{{prefix|}}}7.2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|deepskyblue}}};"}}}>3</span>]] | + | |[[{{{prefix|}}}7.3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|deepskyblue}}};"}}}>3</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|red}}};"}}}>4</span>]] | + | |[[{{{prefix|}}}7.4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|red}}};"}}}>4</span>]] |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]] | + | |[[{{{prefix|}}}7.5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]] |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] | + | |[[{{{prefix|}}}7.6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]] | + | |[[{{{prefix|}}}7.7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]] |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]] | + | |[[{{{prefix|}}}7.8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]] |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] | + | |[[{{{prefix|}}}7.9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|deepskyblue}}};"}}}>10</span>]] | + | |[[{{{prefix|}}}7.10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|deepskyblue}}};"}}}>10</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|red}}};"}}}>11</span>]] | + | |[[{{{prefix|}}}7.11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|red}}};"}}}>11</span>]] |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]] | + | |[[{{{prefix|}}}7.12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]] |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] | + | |[[{{{prefix|}}}7.13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]] | + | |[[{{{prefix|}}}7.14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]] |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]] | + | |[[{{{prefix|}}}7.15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]] |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] | + | |[[{{{prefix|}}}7.16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|deepskyblue}}};"}}}>17</span>]] | + | |[[{{{prefix|}}}7.17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|deepskyblue}}};"}}}>17</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|red}}};"}}}>18</span>]] | + | |[[{{{prefix|}}}7.18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|red}}};"}}}>18</span>]] |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]] | + | |[[{{{prefix|}}}7.19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]] |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]] | + | |[[{{{prefix|}}}7.20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]] |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>]] | + | |[[{{{prefix|}}}7.21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|inherit}}};"}}}>21</span>]] |
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]] | + | |[[{{{prefix|}}}7.22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|inherit}}};"}}}>22</span>]] |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]] | + | |[[{{{prefix|}}}7.23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]] |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|deepskyblue}}};"}}}>24</span>]] | + | |[[{{{prefix|}}}7.24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|deepskyblue}}};"}}}>24</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|red}}};"}}}>25</span>]] | + | |[[{{{prefix|}}}7.25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|red}}};"}}}>25</span>]] |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]] | + | |[[{{{prefix|}}}7.26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]] |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]] | + | |[[{{{prefix|}}}7.27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]] |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>]] | + | |[[{{{prefix|}}}7.28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|inherit}}};"}}}>28</span>]] |
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]] | + | |[[{{{prefix|}}}7.29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|inherit}}};"}}}>29</span>]] |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]] | + | |[[{{{prefix|}}}7.30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]] |
| - | |[[{{{prefix|}}}31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}>31</span>]] | + | |[[{{{prefix|}}}7.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|deepskyblue}}};"}}}>31</span>]] |
|- | |- | ||
|- style="background:{{{color3|#eeeeff}}};" | |- style="background:{{{color3|#eeeeff}}};" | ||
| Line 65: | Line 65: | ||
|width="14%"|[[S|<span style="color:deepskyblue;">S</span>]] | |width="14%"|[[S|<span style="color:deepskyblue;">S</span>]] | ||
|- | |- | ||
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|red}}};"}}}>1</span>]] | + | |[[{{{prefix|}}}8.1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|red}}};"}}}>1</span>]] |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] | + | |[[{{{prefix|}}}8.2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]] | + | |[[{{{prefix|}}}8.3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]] |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span>]] | + | |[[{{{prefix|}}}8.4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|inherit}}};"}}}>4</span>]] |
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]] | + | |[[{{{prefix|}}}8.5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|inherit}}};"}}}>5</span>]] |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] | + | |[[{{{prefix|}}}8.6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|deepskyblue}}};"}}}>7</span>]] | + | |[[{{{prefix|}}}8.7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|deepskyblue}}};"}}}>7</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|red}}};"}}}>8</span>]] | + | |[[{{{prefix|}}}8.8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|red}}};"}}}>8</span>]] |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] | + | |[[{{{prefix|}}}8.9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]] | + | |[[{{{prefix|}}}8.10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]] |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span>]] | + | |[[{{{prefix|}}}8.11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|inherit}}};"}}}>11</span>]] |
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]] | + | |[[{{{prefix|}}}8.12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|inherit}}};"}}}>12</span>]] |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] | + | |[[{{{prefix|}}}8.13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|deepskyblue}}};"}}}>14</span>]] | + | |[[{{{prefix|}}}8.14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|deepskyblue}}};"}}}>14</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span>]] | + | |[[{{{prefix|}}}8.15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|red}}};"}}}>15</span>]] |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] | + | |[[{{{prefix|}}}8.16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]] | + | |[[{{{prefix|}}}8.17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]] |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span>]] | + | |[[{{{prefix|}}}8.18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|inherit}}};"}}}>18</span>]] |
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]] | + | |[[{{{prefix|}}}8.19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|inherit}}};"}}}>19</span>]] |
| - | |[[{{{prefix|}}}20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]] | + | |[[{{{prefix|}}}8.20{{{suffix|}}}|<span style={{{s20|"color:{{{c20|inherit}}};"}}}>20</span>]] |
| - | |[[{{{prefix|}}}21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|deepskyblue}}};"}}}>21</span>]] | + | |[[{{{prefix|}}}8.21{{{suffix|}}}|<span style={{{s21|"color:{{{c21|deepskyblue}}};"}}}>21</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|red}}};"}}}>22</span>]] | + | |[[{{{prefix|}}}8.22{{{suffix|}}}|<span style={{{s22|"color:{{{c22|red}}};"}}}>22</span>]] |
| - | |[[{{{prefix|}}}23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]] | + | |[[{{{prefix|}}}8.23{{{suffix|}}}|<span style={{{s23|"color:{{{c23|inherit}}};"}}}>23</span>]] |
| - | |[[{{{prefix|}}}24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span>]] | + | |[[{{{prefix|}}}8.24{{{suffix|}}}|<span style={{{s24|"color:{{{c24|inherit}}};"}}}>24</span>]] |
| - | |[[{{{prefix|}}}25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span>]] | + | |[[{{{prefix|}}}8.25{{{suffix|}}}|<span style={{{s25|"color:{{{c25|inherit}}};"}}}>25</span>]] |
| - | |[[{{{prefix|}}}26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]] | + | |[[{{{prefix|}}}8.26{{{suffix|}}}|<span style={{{s26|"color:{{{c26|inherit}}};"}}}>26</span>]] |
| - | |[[{{{prefix|}}}27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]] | + | |[[{{{prefix|}}}8.27{{{suffix|}}}|<span style={{{s27|"color:{{{c27|inherit}}};"}}}>27</span>]] |
| - | |[[{{{prefix|}}}28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|deepskyblue}}};"}}}>28</span>]] | + | |[[{{{prefix|}}}8.28{{{suffix|}}}|<span style={{{s28|"color:{{{c28|deepskyblue}}};"}}}>28</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|red}}};"}}}>29</span>]] | + | |[[{{{prefix|}}}8.29{{{suffix|}}}|<span style={{{s29|"color:{{{c29|red}}};"}}}>29</span>]] |
| - | |[[{{{prefix|}}}30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]] | + | |[[{{{prefix|}}}8.30{{{suffix|}}}|<span style={{{s30|"color:{{{c30|inherit}}};"}}}>30</span>]] |
| - | |[[{{{prefix|}}}31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|inherit}}};"}}}>31</span>]] | + | |[[{{{prefix|}}}8.31{{{suffix|}}}|<span style={{{s31|"color:{{{c31|inherit}}};"}}}>31</span>]] |
|colspan="4"| | |colspan="4"| | ||
|- style="background:{{{color3|#eeeeff}}};" | |- style="background:{{{color3|#eeeeff}}};" | ||
| Line 118: | Line 118: | ||
|- | |- | ||
|colspan ="3"| | |colspan ="3"| | ||
| - | |[[{{{prefix|}}}1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]] | + | |[[{{{prefix|}}}9.1{{{suffix|}}}|<span style={{{s1|"color:{{{c1|inherit}}};"}}}>1</span>]] |
| - | |[[{{{prefix|}}}2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] | + | |[[{{{prefix|}}}9.2{{{suffix|}}}|<span style={{{s2|"color:{{{c2|inherit}}};"}}}>2</span>]] |
| - | |[[{{{prefix|}}}3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]] | + | |[[{{{prefix|}}}9.3{{{suffix|}}}|<span style={{{s3|"color:{{{c3|inherit}}};"}}}>3</span>]] |
| - | |[[{{{prefix|}}}4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|deepskyblue}}};"}}}>4</span>]] | + | |[[{{{prefix|}}}9.4{{{suffix|}}}|<span style={{{s4|"color:{{{c4|deepskyblue}}};"}}}>4</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|red}}};"}}}>5</span>]] | + | |[[{{{prefix|}}}9.5{{{suffix|}}}|<span style={{{s5|"color:{{{c5|red}}};"}}}>5</span>]] |
| - | |[[{{{prefix|}}}6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] | + | |[[{{{prefix|}}}9.6{{{suffix|}}}|<span style={{{s6|"color:{{{c6|inherit}}};"}}}>6</span>]] |
| - | |[[{{{prefix|}}}7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]] | + | |[[{{{prefix|}}}9.7{{{suffix|}}}|<span style={{{s7|"color:{{{c7|inherit}}};"}}}>7</span>]] |
| - | |[[{{{prefix|}}}8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]] | + | |[[{{{prefix|}}}9.8{{{suffix|}}}|<span style={{{s8|"color:{{{c8|inherit}}};"}}}>8</span>]] |
| - | |[[{{{prefix|}}}9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] | + | |[[{{{prefix|}}}9.9{{{suffix|}}}|<span style={{{s9|"color:{{{c9|inherit}}};"}}}>9</span>]] |
| - | |[[{{{prefix|}}}10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]] | + | |[[{{{prefix|}}}9.10{{{suffix|}}}|<span style={{{s10|"color:{{{c10|inherit}}};"}}}>10</span>]] |
| - | |[[{{{prefix|}}}11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}>11</span>]] | + | |[[{{{prefix|}}}9.11{{{suffix|}}}|<span style={{{s11|"color:{{{c11|deepskyblue}}};"}}}>11</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|red}}};"}}}>12</span>]] | + | |[[{{{prefix|}}}9.12{{{suffix|}}}|<span style={{{s12|"color:{{{c12|red}}};"}}}>12</span>]] |
| - | |[[{{{prefix|}}}13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] | + | |[[{{{prefix|}}}9.13{{{suffix|}}}|<span style={{{s13|"color:{{{c13|inherit}}};"}}}>13</span>]] |
| - | |[[{{{prefix|}}}14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]] | + | |[[{{{prefix|}}}9.14{{{suffix|}}}|<span style={{{s14|"color:{{{c14|inherit}}};"}}}>14</span>]] |
| - | |[[{{{prefix|}}}15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]] | + | |[[{{{prefix|}}}9.15{{{suffix|}}}|<span style={{{s15|"color:{{{c15|inherit}}};"}}}>15</span>]] |
| - | |[[{{{prefix|}}}16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] | + | |[[{{{prefix|}}}9.16{{{suffix|}}}|<span style={{{s16|"color:{{{c16|inherit}}};"}}}>16</span>]] |
| - | |[[{{{prefix|}}}17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]] | + | |[[{{{prefix|}}}9.17{{{suffix|}}}|<span style={{{s17|"color:{{{c17|inherit}}};"}}}>17</span>]] |
| - | |[[{{{prefix|}}}18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}>18</span>]] | + | |[[{{{prefix|}}}9.18{{{suffix|}}}|<span style={{{s18|"color:{{{c18|deepskyblue}}};"}}}>18</span>]] |
|- | |- | ||
| - | |[[{{{prefix|}}}19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|red}}};"}}}>19</span>]] | + | |[[{{{prefix|}}}9.19{{{suffix|}}}|<span style={{{s19|"color:{{{c19|red}}};"}}}>19</span>]] |
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Revision as of 10:47, 6 October 2010
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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)
Attendance: Nakamura, Kakuda, Saka, Yasumoto, Teoh
- Competent cells preparation (continued)
- Preparation of glucose solution for making SOC medium.
- Inoculation of pre-culture SOB from o/n-incubated agar plate colonies.
- (Night) Transfer from pre-culture to growth culture.
August 4 (Wed)
Attendance: Nakamura, Saka, Kakuda, Yasumoto, Torigata, Teoh
- 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 |
| 2-20H | <bbpart>BBa_K118022</bbpart> | A | C. fermi exocellulase Cex coding |
| 1-2M | <bbpart>BBa_B0034</bbpart> | A | RBS |
| 1-13D | <bbpart>BBa_B0010</bbpart> | A | terminator |
| 1-1D | <bbpart>BBa_R0010</bbpart> | A | promoter |
| 1-18F | <bbpart>BBa_E1010</bbpart> | K | RFP coding |
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.
August 6 (Fri)
Attendance: Nakamura, Saka, Yasumoto, Takino, Teoh
- Colony check
- All transformed cells produced colonies!
- 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>) |
| 1-3A | <bbpart>pSB1C3</bbpart> | C | (" ") |
| 1-5A | <bbpart>pSB1K3</bbpart> | K | (" ") |
- Meeting
August 8 (Sun)
Attendance: Nakamura, Yasumoto
- Colony check
- All parts successfully transformed
- 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
- 1st run: 1-3A, 1-5A (newly miniprepped), 1-1D, 1-18F, 1-13D, 1-2M (repeat)
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 |
| 1-2J | <bbpart>BBa_J32015</bbpart> | A,K | PelB leader sequence |
August 18 (Wed)
iGEM Japan Meet-Up in Kyoto Attendance: Nakamura, Yasumoto, Saka, Kakuda
August 19 (Thu)
- 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 |
| F1 | N/A | A | beta-galactosidase from Edinburgh team |
| F2 | N/A | C | RBS + F1 |
| F3 | N/A | C | Lac promoter + RBS + F1 |
- O/N incubation at 37℃ as per normal
August 23 (Mon)
- Transfer of 2-4A, F1, F2, F3 transformed yesterday to solution culture
- Miniprep & 'Cut check' of <1>
- cut at EcoRI, SpeI
- 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 |
| 3-11I | <bbpart>K105027</bbpart> | A | 'cyc100' minimal promoter |
August 24 (Tue)
- Colony check of part transformed yesterday: both 2-2O & 3-11I produced >100 colonies
- transfer to solution culture
- Miniprep of 2-4A, F1, F2, F3 followed by 'cut check' with EcoRI, SpeI
- (RESULTS)
- Transfer of F1 to solution culture (why?)
- Preparation of glycerol stock of cell culture containing <1> (why?)
- 200ml of culture solution mixed with 100ml of 50% glycerol
- stored at -80℃
August 25 (Wed)
- Miniprep of parts in solution culture: 2-2O, 3-11I, F1
- Cut check of 3-11I & F1 with EcoRI, SpeI
- (RESULTS?)
