From 2010.igem.org
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| | [[Image:KAIST-exp23.png|700px]] | | [[Image:KAIST-exp23.png|700px]] |
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| | + | <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">F_Fusion antibody receptor</td><td width = "200px">60*C</td><td width = "223px">100%</td><td width = "70px">1095bp</td> |
| | + | </tr> |
| | + | <tr align=center> |
| | + | <td width = "40px">b</td><td width = "255px">F_Fusion antibody receptor</td><td width = "200px">58*C</td><td width = "223px">100%</td><td width = "70px">1095bp</td> |
| | + | </tr> |
| | + | <tr align=center> |
| | + | <td width = "40px">c</td><td width = "255px">Antibody</td><td width = "200px">60*C</td><td width = "223px">100%</td><td width = "70px">669bp</td> |
| | + | </tr> |
| | + | <tr align=center> |
| | + | <td width = "40px">d</td><td width = "255px">Antibody</td><td width = "200px">58*C</td><td width = "223px">100%</td><td width = "70px">669bp</td> |
| | + | </tr> |
| | + | <tr align=center> |
| | + | <td width = "40px">e</td><td width = "255px">VF_Fusion antibody receptor</td><td width = "200px">60*C</td><td width = "223px">100%</td><td width = "70px">1107bp</td> |
| | + | </tr> |
| | + | <tr align=center> |
| | + | <td width = "40px">f</td><td width = "255px">VF_Fusion antibody receptor</td><td width = "200px">58*C</td><td width = "223px">100%</td><td width = "70px">1107bp</td> |
| | + | </tr> |
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| | + | </table> |
| | + | <br> |
| | + | <br> |
| | + | [[Image:KAIST-exp24.png|360px]] |
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Revision as of 19:37, 16 September 2010
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PCR Experiment
9/6 ~ 9/9 PCR Result
PCR composition
| Materials | Amount |
| Template | 100pmole |
| Primer A | 50pmole |
| Primer B | 50pmole |
| 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 |
| Total | 50ul |
Exp 1
| Segment | Annealing Temperature | Result |
| a | STAT1 | 53*C | - | - |
| b | V_STAT1 | 53*C | - | - |
| c | Fusion Antibody receptor | 53*C | - | - |
| d | Signal peptide | 56*C | - | - |
| e | Ig-like | 56*C | - | - |
| f | Signal Peptide + Ig-like | 56*C | - | - |
| g | Antibody | 56*C | - | - |
| h | V_Fusion Antibody Receptor | 53*C | - | - |
| i | B_FGFR1 | 56*C | - | - |
| j | VB_FGFR1 | 56*C | - | - |
Exp 2
| Segment | Annealing Temperature | Result |
| a | STAT1 | 48*C | 60% | 2139bp |
| b | V_STAT1 | 48*C | - | - |
Exp 3
| Segment | Annealing Temperature | Result |
| a | STAT1 | 50*C | 100% | 2139bp |
| b | V_STAT1 | 50*C | - | - |
Exp 4
| Segment | Annealing Temperature | Result |
| a | Fusion Antibody receptor | 50*C | - | - |
| b | Signal peptide | 50*C | - | - |
| c | Ig-like | 50*C | 60% | 339bp |
| d | Signal Peptide + Ig-like | 50*C | - | - |
| e | Antibody | 50*C | - | - |
| f | V_Fusion Antibody Receptor | 50*C | - | - |
9/10 ~ 9/12 PCR Result
PCR composition
| Materials | Amount |
| Template | 150pmole |
| Primer A | 50pmole |
| Primer B | 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 |
| Total | 50ul |
Exp 1
| Segment | Annealing Temperature | Result |
| a | STAT1 | 60*C | - | - |
| b | V_STAT1 | 60*C | - | - |
| c | Fusion Antibody receptor | 60*C | 70% | 1095bp |
| d | Signal peptide | 60*C | - | - |
| e | Ig-like | 60*C | 60% | 339bp |
| f | Signal Peptide + Ig-like | 60*C | - | - |
| g | Antibody | 60*C | 100% | 669bp |
| h | V_Fusion Antibody Receptor | 60*C | - | - |
| i | B_FGFR1 | 60*C | - | - |
| j | VB_FGFR1 | 60*C | - | - |
| x1 | negative 1 | 60*C | - | - |
| x2 | negative 2 | 60*C | - | - |
Exp 2
| Segment | Annealing Temperature | Result |
| a | STAT1 | 58*C | 60% | 2139bp |
| b | V_STAT1 | 58*C | - | - |
| c | Fusion Antibody receptor | 58*C | 80% | 1095bp |
| d | Signal peptide | 58*C | 80% | 87bp |
| e | Ig-like | 58*C | 100% | 339bp |
| f | Signal Peptide + Ig-like | 58*C | 80% | 426bp |
| g | Antibody | 58*C | 100% | 669bp |
| h | V_Fusion Antibody Receptor | 58*C | - | - |
| i | B_FGFR1 | 58*C | - | - |
| j | VB_FGFR1 | 58*C | - | - |
| x | negative | 58*C | - | - |
Exp 3
| Segment | Annealing Temperature | Result |
| a | STAT1 | 62*C | - | - |
| b | V_STAT1 | 62*C | - | - |
| c | Fusion Antibody receptor | 62*C | 10% | 1095bp |
| d | Signal peptide | 62*C | - | - |
| e | Ig-like | 62*C | - | - |
| f | Signal Peptide + Ig-like | 62*C | - | - |
| g | Antibody | 62*C | - | - |
| h | V_Fusion Antibody Receptor | 62*C | - | - |
| i | B_FGFR1 | 62*C | - | - |
| j | VB_FGFR1 | 62*C | - | - |
| x | negative | 62*C | - | - |
Exp 4
| Segment | Annealing Temperature | Result |
| a | STAT1 | 56*C | - | - |
| b | V_STAT1 | 56*C | - | - |
| c | Fusion Antibody receptor | 56*C | 5% | 1095bp |
| d | Signal peptide | 56*C | - | - |
| e | Ig-like | 56*C | 80% | 339bp |
| f | Signal Peptide + Ig-like | 56*C | - | - |
| g | Antibody | 56*C | 80% | 669bp |
| h | V_Fusion Antibody Receptor | 56*C | - | - |
| i | B_FGFR1 | 56*C | - | - |
| j | VB_FGFR1 | 56*C | - | - |
| x | negative | 56*C | - | - |
Exp 5
| Segment | Annealing Temperature | Result |
| a | V_STAT1 | 64*C | - | - |
| b | B_FGFR1 | 64*C | 5% | 1398bp |
| c | VB_FGFR1 | 64*C | - | - |
Exp 6
| Segment | Annealing Temperature | Result |
| a | V_STAT1 | 64*C | - | - |
| b | B_FGFR1 | 64*C | 3% | 1398bp |
| c | VB_FGFR1 | 64*C | - | - |
Exp 7
| Segment | Annealing Temperature | Result |
| a | 3'UTR | 62*C | - | - |
| b | 5'UTR | 62*C | - | - |
Exp 8
| Segment | Annealing Temperature | Result |
| a | V_STAT1 | 68*C | - | - |
| b | B_FGFR1 | 68*C | 10% | 1398bp |
| c | VB_FGFR1 | 68*C | - | - |
Exp 9
| Segment | Annealing Temperature | Result |
| a | V_STAT1 | 70*C | - | - |
| b | B_FGFR1 | 70*C | 3% | 1398bp |
| c | VB_FGFR1 | 70*C | - | - |
| c | 5'UTR | 70*C | - | - |
| c | 3'UTR | 70*C | - | - |
Exp 10
| Segment | Annealing Temperature | Result |
| a | V_STAT1 | 67*C | - | - |
| b | B_FGFR1 | 67*C | 6% | 1398bp |
| c | VB_FGFR1 | 67*C | - | - |
9/13 ~ 9/16 PCR Result
PCR composition
| Materials | Amount |
| Template | 100pmole |
| Primer A | 50pmole |
| Primer B | 50pmole |
| Bioneer Accupower HF PCR premix | < 1ul |
| DW (3rd sterile water) | rest |
| Total | 50ul |
Exp 1
| Segment | Annealing Temperature | Result |
| a | STAT1 | 50*C | 100% | 2139bp |
| b | STAT1 | 52*C | 100% | 2139bp |
| c | STAT1 | 54*C | 100% | 2139bp |
| d | STAT1 | 56*C | 100% | 2139bp |
| e | STAT1 | 58*C | 100% | 2139bp |
| f | STAT1 | 60*C | 100% | 2139bp |
| g | V_STAT1 | 50*C | 100% | 2151bp |
| h | V_STAT1 | 52*C | 100% | 2151bp |
| i | V_STAT1 | 54*C | 100% | 2151bp |
| j | V_STAT1 | 56*C | 100% | 2151bp |
| k | V_STAT1 | 58*C | 100% | 2151bp |
| l | V_STAT1 | 60*C | 100% | 2151bp |
Exp 2
| Segment | Annealing Temperature | Result |
| a | B_FGFR | 50*C | 100% | 1398bp |
| b | B_FGFR | 52*C | 100% | 1398bp |
| c | B_FGFR | 54*C | 100% | 1398bp |
| d | B_FGFR | 56*C | 100% | 1398bp |
| e | B_FGFR | 58*C | 100% | 1398bp |
| f | B_FGFR | 60*C | 100% | 1398bp |
| g | VB_FGFR | 50*C | 100% | 1420bp |
| h | VB_FGFR | 52*C | 100% | 1420bp |
| i | VB_FGFR | 54*C | 100% | 1420bp |
| j | VB_FGFR | 56*C | 100% | 1420bp |
| k | VB_FGFR | 58*C | 100% | 1420bp |
| l | VB_FGFR | 60*C | 100% | 1420bp |
Exp 3
| Segment | Annealing Temperature | Result |
| a | Signal peptide | 61*C | 30% | 87bp |
| b | Ig-like | 61*C | 100% | 339bp |
| c | Signal peptide + Ig-like | 61*C | 100% | 426bp |
d | Signal peptide | 59*C | 30% | 87bp |
</tr>
| e | Ig-like | 59*C | 100% | 339bp |
| f | Signal peptide + Ig-like | 59*C | 100% | 426bp |
| g | 3'UTR | 52*C | 50% | 444bp |
| h | 3'UTR | 54*C | 50% | 444bp |
| i | 3'UTR | 56*C | 100% | 444bp |
| j | 3'UTR | 58*C | 100% | 444bp |
| k | 3'UTR | 60*C | 100% | 444bp |
| l | 5'UTR | 52*C | 100% | 500bp |
| m | 5'UTR | 54*C | 100% | 500bp |
| n | 5'UTR | 56*C | 100% | 500bp |
| o | 5'UTR | 58*C | 100% | 500bp |
| p | 5'UTR | 69*C | 70% | 500bp |
Exp 4
| Segment | Annealing Temperature | Result |
| a | F_Fusion antibody receptor | 60*C | 100% | 1095bp |
| b | F_Fusion antibody receptor | 58*C | 100% | 1095bp |
| c | Antibody | 60*C | 100% | 669bp |
| d | Antibody | 58*C | 100% | 669bp |
| e | VF_Fusion antibody receptor | 60*C | 100% | 1107bp |
| f | VF_Fusion antibody receptor | 58*C | 100% | 1107bp |
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