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Team:KAIST-Korea/Project/Methods/PCR
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| + | <a href="https://2010.igem.org/Team:KAIST-Korea/Project/Methods"><img src="https://static.igem.org/mediawiki/2010/6/6f/Method_button.png" width = 300></a> | ||
| + | <a href="https://2010.igem.org/Team:KAIST-Korea/Project/Methods/PCR"><img src="https://static.igem.org/mediawiki/2010/e/e2/PCR-KAIST3.png" width = 300></a> | ||
| + | <a href="https://2010.igem.org/Team:KAIST-Korea/Project/Methods/PCR_Result"><img src="https://static.igem.org/mediawiki/2010/9/94/PCR-Exp.png" width = 300></a></center> | ||
| + | </html> | ||
| + | <br> | ||
| + | <br> | ||
<span style=font-size:30px><font face="Maiandra GD"><b>PCR Strategy</b></font></span> | <span style=font-size:30px><font face="Maiandra GD"><b>PCR Strategy</b></font></span> | ||
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| - | [[Image: | + | [[Image:PCR_STAT1_m.jpg]] |
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<td>PC_LINKR</td><td>50pmole</td> | <td>PC_LINKR</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> | ||
| + | |||
| + | [[Image:PCR_cycle.jpg]] | ||
| + | </center> | ||
| + | <br> | ||
| + | <br> | ||
| + | <br> | ||
| + | |||
| + | === GAS Promoter (expression) === | ||
| + | |||
| + | <center> | ||
| + | [[Image:PCR_GAS.jpg]] | ||
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| + | <br> | ||
| + | |||
| + | <table border=1> | ||
| + | <tr align=center> | ||
| + | <td><b>Primer</b></td><td><b>Direction</b></td><td><b>Sequences</b></td><td><b>Length</b></td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>PP_F (Proximal promoter)</td><td>Forward</td><td>5’-<font color = blue>CGC</font><font color = red>CTGCAG</font><font color = blue>GCG</font>AATAGCTGG–3’</td><td>21bp</td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>CGFP_R (Core promoter + GFP)</td><td>Reverse</td><td>3’-CGACATATGCTTGTACAGCTCG-5’</td><td>22bp</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <table border=1> | ||
| + | <tr align=center> | ||
| + | <td><b>Materials</b></td><td><b>Amount</b></td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>Proximal promoter(template)</td><td>150pmole</td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>Core promoter + GFP(template)</td><td>150pmole</td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>PP_F (Proximal promoter)</td><td>50pmole</td> | ||
| + | </tr> | ||
| + | <tr align=center> | ||
| + | <td>CGFP_R (Core promoter + GFP)</td><td>50pmole</td> | ||
</tr> | </tr> | ||
<tr align=center> | <tr align=center> | ||
Latest revision as of 07:08, 20 October 2010
|
 PCR Strategy
V_STAT1 (expression)
STAT1 (submission)
Fusion Antibody Receptor (Submission)
Signal Peptide (Submission)
Ig-like (Submission)
Signal Peptide + Ig-like (Submission)
Antibody (Submission)
V_Fusion Antibody Receptor (Expression)
B_FGFR1 (Submission)
VB_FGFR1 (Expression)
Fusion Antibody Receptor (Expression)
5’UTR (expression)
3’UTR (expression)
Selection Marker (expression)
Core promoter + GFP (expression)
Proximal promoter (expression)
GAS Promoter (expression)
HA1 (expression)
HA2 (expression)
Fusion antibody receptor (expression)
5’UTR + Fusion Antibody receptor (expression)
HA1 + Selection Marker (expression)
HA2 + 3'UTR (expression)
5’UTR - Fusion Antibody receptor + HA1 - Selection Marker (expression)
Homologous Recombinant (expression)
|
