Team:KAIST-Korea/Notebook/Diary/September

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(Difference between revisions)
(→September next)
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<br>
<br>
-
== September next ==
+
==9/6 ~ 9/9 PCR==
-
 
+
 +
<span style=font-size:12px><b>PCR composition </b></span>
 +
<table border=1>
 +
<tr align=center>
 +
<td><b>Materials</b></td><td><b>Amount</b></td>
</tr>
</tr>
 +
<tr align=center>
 +
<td>Template</td><td>100pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Primer A</td><td>50pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Primer B</td><td>50pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>dNTP (dNTP mixture 2.5mM TaKaRa)</td><td>2ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)</td><td>0.25ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>10x buffer (10x cloned Pfu Reaction buffer)</td><td>5ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>DW (3rd sterile water)</td><td>rest</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Total</td><td>50ul</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
<b>Exp 1</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">53*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">53*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">Fusion Antibody receptor</td><td width = "200px">53*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px">Signal peptide</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px">Ig-like</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px">Signal Peptide + Ig-like</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">g</td><td width = "255px">Antibody</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">53*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp1.png|560px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 2</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">48*C</td><td width = "223px">60%</td><td width = "70px">2139bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">48*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp2.png|260px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 3</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px"><font color = red>STAT1</font></td><td width = "200px">50*C</td><td width = "223px">100%</td><td width = "70px">2139bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp3.png|260px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 4</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">Fusion Antibody receptor</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">Signal peptide</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">Ig-like</td><td width = "200px">50*C</td><td width = "223px">60%</td><td width = "70px">339bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px">Signal Peptide + Ig-like</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px">Antibody</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">50*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp4.png|300px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
==9/10 ~ 9/12 PCR==
 +
 +
<span style=font-size:12px><b>PCR composition </b></span>
 +
<table border=1>
 +
<tr align=center>
 +
<td><b>Materials</b></td><td><b>Amount</b></td>
 +
</tr>
 +
<tr align=center>
 +
<td>Template</td><td>150pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Primer A</td><td>50pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Primer B</td><td>50pmole</td>
 +
</tr>
 +
<tr align=center>
 +
<td>dNTP (dNTP mixture 2.5mM TaKaRa)</td><td>5ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)</td><td>0.25ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>10x buffer (10x cloned Pfu Reaction buffer)</td><td>5ul</td>
 +
</tr>
 +
<tr align=center>
 +
<td>DW (3rd sterile water)</td><td>rest</td>
 +
</tr>
 +
<tr align=center>
 +
<td>Total</td><td>50ul</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
<b>Exp 1</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">Fusion Antibody receptor</td><td width = "200px">60*C</td><td width = "223px">70%</td><td width = "70px">1095bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px">Signal peptide</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px">Ig-like</td><td width = "200px">60*C</td><td width = "223px">60%</td><td width = "70px">339bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px">Signal Peptide + Ig-like</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">g</td><td width = "255px"><font color = red>Antibody</font></td><td width = "200px">60*C</td><td width = "223px">100%</td><td width = "70px">669bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">x1</td><td width = "255px">negative 1</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">x2</td><td width = "255px">negative 2</td><td width = "200px">60*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp5.png|700px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 2</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">58*C</td><td width = "223px">60%</td><td width = "70px">2139bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">58*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px"><font color = blue>Fusion Antibody receptor</font></td><td width = "200px">58*C</td><td width = "223px">80%</td><td width = "70px">1095bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px"><font color = blue>Signal peptide</font></td><td width = "200px">58*C</td><td width = "223px">80%</td><td width = "70px">87bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px"><font color = red>Ig-like</font></td><td width = "200px">58*C</td><td width = "223px">100%</td><td width = "70px">339bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px"><font color = blue>Signal Peptide + Ig-like</font></td><td width = "200px">58*C</td><td width = "223px">80%</td><td width = "70px">426bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">g</td><td width = "255px"><font color = red>Antibody</font></td><td width = "200px">58*C</td><td width = "223px">100%</td><td width = "70px">669bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">58*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">58*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">58*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">x</td><td width = "255px">negative</td><td width = "200px">58*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp6.png|700px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 3</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">Fusion Antibody receptor</td><td width = "200px">62*C</td><td width = "223px">10%</td><td width = "70px">1095bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px">Signal peptide</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px"><font color = red>Ig-like</font></td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px">Signal Peptide + Ig-like</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">g</td><td width = "255px">Antibody<td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">x</td><td width = "255px">negative</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp7.png|640px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 4</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">STAT1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">V_STAT1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">Fusion Antibody receptor</td><td width = "200px">56*C</td><td width = "223px">5%</td><td width = "70px">1095bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">d</td><td width = "255px">Signal peptide</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">e</td><td width = "255px"><font color = blue>Ig-like</font></td><td width = "200px">56*C</td><td width = "223px">80%</td><td width = "70px">339bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">f</td><td width = "255px">Signal Peptide + Ig-like</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">g</td><td width = "255px"><font color = blue>Antibody</font></td><td width = "200px">56*C</td><td width = "223px">80%</td><td width = "70px">669bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">x</td><td width = "255px">negative</td><td width = "200px">56*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp8.png|640px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 5</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">V_STAT1</td><td width = "200px">64*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">B_FGFR1</td><td width = "200px">64*C</td><td width = "223px">5%</td><td width = "70px">1398bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">VB_FGFR1</td><td width = "200px">64*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp9.png|320px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 6</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">V_STAT1</td><td width = "200px">64*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">B_FGFR1</td><td width = "200px">64*C</td><td width = "223px">3%</td><td width = "70px">1398bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">VB_FGFR1</td><td width = "200px">64*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp10.png|320px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 7</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">3'UTR</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">5'UTR</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td></tr>
 +
</table>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 8</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">V_STAT1</td><td width = "200px">68*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">B_FGFR1</td><td width = "200px">68*C</td><td width = "223px">10%</td><td width = "70px">1398bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">VB_FGFR1</td><td width = "200px">68*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp12.png|400px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 9</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">V_STAT1</td><td width = "200px">70*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">B_FGFR1</td><td width = "200px">70*C</td><td width = "223px">3%</td><td width = "70px">1398bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">VB_FGFR1</td><td width = "200px">70*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">5'UTR</td><td width = "200px">70*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">3'UTR</td><td width = "200px">70*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp13.png|400px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<b>Exp 10</b>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "300px"><b>Segment</b></td><td width = "200px"><b>Annealing Temperature</b></td><td width = "300px"><b>Result</b></td>
 +
</tr>
 +
</table>
 +
<table border=1>
 +
<tr align=center>
 +
<td width = "40px">a</td><td width = "255px">V_STAT1</td><td width = "200px">67*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">b</td><td width = "255px">B_FGFR1</td><td width = "200px">67*C</td><td width = "223px">6%</td><td width = "70px">1398bp</td>
 +
</tr>
 +
<tr align=center>
 +
<td width = "40px">c</td><td width = "255px">VB_FGFR1</td><td width = "200px">67*C</td><td width = "223px">-</td><td width = "70px">-</td>
 +
</tr>
</table>
</table>
 +
<br>
 +
<br>
 +
[[Image:KAIST-exp14.png|320px]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>
 +
<br>

Revision as of 16:45, 12 September 2010

 

September 1

pREP41 vector has arrived.

-> we will do E.coli transformation and increase its number.

pREP42-GFP vector has arrived (in E.coli)

-> will incubate a day and do spreading for further experiment.

Making culture media

  1. 500mL LB broth + agar 1.5% (7.5g)
  2. Do auto clave
  3. Cool it down until it reaches 50~60℃
  4. Add ampicillin(1000x 100mg/mL) and stir
  5. putting out bubbles on culture media
  6. pouling on petri dish and wait for 1 hour

Culture

  1. Competent cell + DN10A and leave it in ice for 30min
  2. Do heat shock for 45 sec and store it in ice for 2min
  3. Stabilize it in incubator for 30 to 40min
  4. Do spreading


September ?

V_STAT1 (expression)

PCR STAT1.jpg


PrimerDirectionSequencesLength
V_STAT_FForward(NdeI)5’-TGTTCATATGGCTAATGTCTCAGTGGTACGAACTTCAG-3’24bp
V_STAT_RReverse(BamHI)5’-TATCGGATCCGAATTTACACTTCAGACACAGAAATCAAC-3’25bp


MaterialsAmount
STAT1(KRIBB) (template)150pmole
V_STAT_F50pmole
V_STAT_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




STAT1 (submission)

PCR STAT1.jpg


PrimerDirectionSequencesLength
STAT_FForward5’-ATGTCTCAGTGGTACGAACTTCAG-3’24bp
STAT_RReverse5’-TTACACTTCAGACACAGAAATCAAC-3’25bp


MaterialsAmount
STAT1(KRIBB) (template)150pmole
STAT_F50pmole
STAT_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Fusion Antibody Receptor (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
SIG_FForward5’-ATGGTCTTTTTAAATTCCTCTCCC-3’24bp
Ab_RReverse5’-GGGGCTGTTGTTTTGGCTGAGG-3’22bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
SIG_F50pmole
Ab_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Signal Peptide (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
SIG_FForward5’-ATGGTCTTTTTAAATTCCTCTCCC-3’24bp
SIG_RReverse5’-AGCCGCCACCAACCGAGTAGAAA-3’23bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
SIG_F50pmole
SIG_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Ig-like (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
IG_FForward5’-AGGCCGTCCCCGACCTTGCCTG-3’22bp
IG_RReverse5’-GGAGTCATCATCATCATCATCATC-3’24bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
IG_F50pmole
IG_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Signal Peptide + Ig-like (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
SIG_FForward5’-ATGGTCTTTTTAAATTCCTCTCCC-3’24bp
IG_RReverse5’-GGAGTCATCATCATCATCATCATC-3’24bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
SIG_F50pmole
IG_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Antibody (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
Ab_FForward5’-ACCCAGTCTCCAGCAATCATGTC-3’23bp
Ab_RReverse5’-GGGGCTGTTGTTTTGGCTGAGG-3’22bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
Ab_F50pmole
Ab_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




V_Fusion Antibody Receptor (Expression)

PCR FGFR.jpg


PrimerDirectionSequencesLength
SIG_FForward5’-ATGGTCTTTTTAAATTCCTCTCCC-3’24bp
Linker_RReverse5’-CTCTCTTCCAGGGCTTCCAGAACGGGGCTGTTGTTTTGGCTGAGGA-3’46(23+23)bp


MaterialsAmount
Fusion antibody receptor T vector (template)150pmole
SIG_F50pmole
Linker_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




B_FGFR1 (Submission)

PCR FGFR.jpg


PrimerDirectionSequencesLength
B_FGFR_FForward5’-GTTCTGGAAGCCCTGGAAGAGAG-3’23bp
B_FGFR_RReverse5’-TCAGCGGCGTTTGAGTCCGCCAT-3’23bp


MaterialsAmount
FGFR1(KRIBB) (template)150pmole
B_FGFR_F50pmole
B_FGFR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




VB_FGFR1 (Expression)

PCR FGFR.jpg


PrimerDirectionSequencesLength
Linker_FForward5’-TCCTCAGCCAAAACAACAGCCCCGTTCTGGAAGCCCTGGAAGAGAG-3’46(23+23)bp
B_FGFR_RReverse5’-TCAGCGGCGTTTGAGTCCGCCAT-3’23bp


MaterialsAmount
FGFR1(KRIBB) (template)150pmole
Linker_F50pmole
B_FGFR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Fusion Antibody Receptor (Expression)

PCR FGFR.jpg


PrimerDirectionSequencesLength
SIG_FForward5’-ATGGTCTTTTTAAATTCCTCTCCC-3’24bp
B_FGFR_RReverse5’-TCAGCGGCGTTTGAGTCCGCCAT-3’23bp


MaterialsAmount
V_Fusion antibody receptor (template)150pmole
VB_FGFR1(2+3) (template)150pmole
SIG_F50pmole
B_FGFR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




5’UTR (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
HR_FForward5’-AAACCAGTTTTGCCAAAACAAC-3’22bp
UF_LinkRReverse5’ –GAGAGGAATTTAAAAAGACCATAAACAACATTTTTCTTTTTTAC - 3’44(22+22)bp


MaterialsAmount
S.pombe genomic DNA (template)150pmole
HR_F50pmole
UF_LinkR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




3’UTR (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
HU_LINKF (HA2 + 3’UTR)Forward5’-CCATATGACCCAGATTACGCTTGGAAATTAAATCGTGCGTAAG-3’43(21+22)bp
HR_RReverse5’ – GCGTTCTTTTTTAATTCTGAATTAA – 3’25bp


MaterialsAmount
S.pombe genomic DNA (template)150pmole
HU_LINKF (HA2 + 3’UTR)50pmole
HR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Selection Marker (expression)

PCR GAS.jpg


PrimerDirectionSequencesLength
HS_LINKF (HA1 + Selection Marker)Forward5’-CCATATGACCCAGATTACGCTAGCAAGCTTAAGCTTAGCTAC-3’42(21+21)bp
SH_LINKRReverse5’-GTCAGGAACATCGTATGGGTAGACCTGCAGAAGCTTAAGCTTG-3’43(21+22)bp


MaterialsAmount
pREP42-GFP vector (template)150pmole
HS_LINKF (HA1 + Selection Marker)50pmole
SH_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Core promoter + GFP (expression)

PCR GAS.jpg


PrimerDirectionSequencesLength
PC_LINKF (Proximal + Core)Forward5’–CAGAATAGACACACGGGCCGACATTGAAGATATATAAAGGAAG–3’43(20+23)bp
CGFP_R (Core promoter + GFP)Reverse3’-CGACATATGCTTGTACAGCTCG-5’22bp


MaterialsAmount
pREP42-GFP vector (template)150pmole
PC_LINKF (Proximal + Core)50pmole
CGFP_R (Core promoter + GFP)50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Proximal promoter (expression)

PCR GAS.jpg


PrimerDirectionSequencesLength
PP_F (Proximal promoter)Forward5’-CGCCTGCAGGCGAATAGCTGG–3’21bp
PC_LINKRReverse5’–CTTCCTTTATATATCTTCAATGTCGGCCCGTGTGTCTATTCTG–3’43(23+20)bp


MaterialsAmount
Proximal promoter T vector (template)150pmole
PP_F (Proximal promoter)50pmole
PC_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




HA1 (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
FH_LINKF (FGFR + HA1)Forward5’-GCGGACTCAAACGCCGCTGATACCCATACGATGTTCCTGAC-3’41(20+21)bp
HS_LINKRReverse5’-GTAGCTAAGCTTAAGCTTGCTAGCGTAATCTGGGTCATATGG-3’42(21+21)bp


MaterialsAmount
HA T vector (template)150pmole
FH_LINKF (FGFR + HA1)50pmole
HS_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




HA2 (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
SH_LINKF (Selection Marker + HA2)Forward5’-CAAGCTTAAGCTTCTGCAGGTCTACCCATACGATGTTCCTGAC-3’43(22+21)bp
HU_LINKRReverse5’-CTTACGCACGATTTAATTTCCAAGCGTAATCTGGGTCATATGG-3’43(22+21)bp


MaterialsAmount
HA T vector (template)150pmole
SH_LINKF (Selection Marker + HA2)50pmole
HU_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Fusion antibody receptor (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
UF_LinkF (5’UTR+FGFR Linker)Forward5’-GTAAAAAAGAAAAATGTTGTTTATGGTCTTTTTAAATTCCTCTC-3’44(22+22)bp
FH_LINKRReverse5’-GTCAGGAACATCGTATGGGTATCAGCGGCGTTTGAGTCCGC-3’41(21+20)bp


MaterialsAmount
Fusion Antibody receptor (template)150pmole
UF_LinkF (5’UTR+FGFR Linker)50pmole
FH_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




5’UTR + Fusion Antibody receptor (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
HR_F (for homologous recombinant)Forward5’-AAACCAGTTTTGCCAAAACAAC-3’22bp
FH_LINKRReverse5’-GTCAGGAACATCGTATGGGTATCAGCGGCGTTTGAGTCCGC-3’41(21+20)bp


MaterialsAmount
5'UTR (template)150pmole
Fusion Antibody receptor (template)150pmole
FH_LINKF (FGFR + HA1)50pmole
SH_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




HA1 + Selection Marker (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
FH_LINKF (FGFR + HA1)Forward5’-GCGGACTCAAACGCCGCTGATACCCATACGATGTTCCTGAC-3’41(20+21)bp
SH_LINKRReverse5’-GTCAGGAACATCGTATGGGTAGACCTGCAGAAGCTTAAGCTTG-3’43(21+22)bp


MaterialsAmount
HA1 (template)150pmole
Selection Marker (template)150pmole
FH_LINKF (FGFR + HA1)50pmole
SH_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




HA2 + 3'UTR (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
SH_LINKF (Selection Marker + HA2)Forward5’-CAAGCTTAAGCTTCTGCAGGTCTACCCATACGATGTTCCTGAC-3’43(22+21)bp
HR_RReverse5’ – GCGTTCTTTTTTAATTCTGAATTAA – 3’25bp


MaterialsAmount
HA2 (template)150pmole
3’UTR (template)150pmole
SH_LINKF (Selection Marker + HA2)50pmole
HR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




5’UTR - Fusion Antibody receptor + HA1 - Selection Marker (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
HR_F(for homologous recombinant)Forward5’-AAACCAGTTTTGCCAAAACAAC-3’22bp
SH_LINKRReverse5’-GTCAGGAACATCGTATGGGTAGACCTGCAGAAGCTTAAGCTTG-3’43(21+22)bp


MaterialsAmount
5’UTR - Fusion Antibody receptor (template) 150pmole
HA1 - Selection Marker (template) 150pmole
HR_F (for homologous recombinant)50pmole
SH_LINKR50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




Homologous Recombinant (expression)

PCR HR.jpg


PrimerDirectionSequencesLength
HR_F(for homologous recombinant)Forward5’-AAACCAGTTTTGCCAAAACAAC-3’22bp
HR_RReverse5’ – GCGTTCTTTTTTAATTCTGAATTAA – 3’25bp


MaterialsAmount
HA2 + 3'UTR (template) 150pmole
5’UTR - Fusion Antibody receptor - HA1 - Selection Marker (template) 150pmole
HR_F (for homologous recombinant)50pmole
HR_R50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul

PCR cycle.jpg




9/6 ~ 9/9 PCR

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)2ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSTAT153*C--
bV_STAT153*C--
cFusion Antibody receptor53*C--
dSignal peptide56*C--
eIg-like56*C--
fSignal Peptide + Ig-like56*C--
gAntibody56*C--
hV_Fusion Antibody Receptor53*C--
iB_FGFR156*C--
jVB_FGFR156*C--



KAIST-exp1.png




Exp 2

SegmentAnnealing TemperatureResult
aSTAT148*C60%2139bp
bV_STAT148*C--



KAIST-exp2.png




Exp 3

SegmentAnnealing TemperatureResult
aSTAT150*C100%2139bp
bV_STAT150*C--



KAIST-exp3.png




Exp 4

SegmentAnnealing TemperatureResult
aFusion Antibody receptor50*C--
bSignal peptide50*C--
cIg-like50*C60%339bp
dSignal Peptide + Ig-like50*C--
eAntibody50*C--
fV_Fusion Antibody Receptor50*C--



KAIST-exp4.png






9/10 ~ 9/12 PCR

PCR composition

MaterialsAmount
Template150pmole
Primer A50pmole
Primer B50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)0.25ul
10x buffer (10x cloned Pfu Reaction buffer)5ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSTAT160*C--
bV_STAT160*C--
cFusion Antibody receptor60*C70%1095bp
dSignal peptide60*C--
eIg-like60*C60%339bp
fSignal Peptide + Ig-like60*C--
gAntibody60*C100%669bp
hV_Fusion Antibody Receptor60*C--
iB_FGFR160*C--
jVB_FGFR160*C--
x1negative 160*C--
x2negative 260*C--



KAIST-exp5.png





Exp 2

SegmentAnnealing TemperatureResult
aSTAT158*C60%2139bp
bV_STAT158*C--
cFusion Antibody receptor58*C80%1095bp
dSignal peptide58*C80%87bp
eIg-like58*C100%339bp
fSignal Peptide + Ig-like58*C80%426bp
gAntibody58*C100%669bp
hV_Fusion Antibody Receptor58*C--
iB_FGFR158*C--
jVB_FGFR158*C--
xnegative58*C--



KAIST-exp6.png




Exp 3

SegmentAnnealing TemperatureResult
aSTAT162*C--
bV_STAT162*C--
cFusion Antibody receptor62*C10%1095bp
dSignal peptide62*C--
eIg-like62*C--
fSignal Peptide + Ig-like62*C--
gAntibody62*C--
hV_Fusion Antibody Receptor62*C--
iB_FGFR162*C--
jVB_FGFR162*C--
xnegative62*C--



KAIST-exp7.png




Exp 4

SegmentAnnealing TemperatureResult
aSTAT156*C--
bV_STAT156*C--
cFusion Antibody receptor56*C5%1095bp
dSignal peptide56*C--
eIg-like56*C80%339bp
fSignal Peptide + Ig-like56*C--
gAntibody56*C80%669bp
hV_Fusion Antibody Receptor56*C--
iB_FGFR156*C--
jVB_FGFR156*C--
xnegative56*C--



KAIST-exp8.png




Exp 5

SegmentAnnealing TemperatureResult
aV_STAT164*C--
bB_FGFR164*C5%1398bp
cVB_FGFR164*C--



KAIST-exp9.png





Exp 6

SegmentAnnealing TemperatureResult
aV_STAT164*C--
bB_FGFR164*C3%1398bp
cVB_FGFR164*C--



KAIST-exp10.png





Exp 7

SegmentAnnealing TemperatureResult
a3'UTR62*C--
b5'UTR62*C--









Exp 8

SegmentAnnealing TemperatureResult
aV_STAT168*C--
bB_FGFR168*C10%1398bp
cVB_FGFR168*C--



KAIST-exp12.png





Exp 9

SegmentAnnealing TemperatureResult
aV_STAT170*C--
bB_FGFR170*C3%1398bp
cVB_FGFR170*C--
c5'UTR70*C--
c3'UTR70*C--



KAIST-exp13.png





Exp 10

SegmentAnnealing TemperatureResult
aV_STAT167*C--
bB_FGFR167*C6%1398bp
cVB_FGFR167*C--



KAIST-exp14.png





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