From 2010.igem.org
(Difference between revisions)
|
|
| Line 54: |
Line 54: |
| | <br> | | <br> |
| | <b>Exp 1</b> | | <b>Exp 1</b> |
| | + | <center> |
| | <table border=1> | | <table border=1> |
| | <tr align=center> | | <tr align=center> |
| Line 91: |
Line 92: |
| | </tr> | | </tr> |
| | </table> | | </table> |
| - | <br>
| |
| | <br> | | <br> |
| | [[Image:KAIST-exp1.png|560px]] | | [[Image:KAIST-exp1.png|560px]] |
| - | <br> | + | </center> |
| - | <br>
| + | |
| - | <br>
| + | |
| | <br> | | <br> |
| | <br> | | <br> |
| | <b>Exp 2</b> | | <b>Exp 2</b> |
| | + | <center> |
| | <table border=1> | | <table border=1> |
| | <tr align=center> | | <tr align=center> |
| Line 113: |
Line 112: |
| | </tr> | | </tr> |
| | </table> | | </table> |
| - | <br>
| |
| | <br> | | <br> |
| | [[Image:KAIST-exp2.png|260px]] | | [[Image:KAIST-exp2.png|260px]] |
| - | <br> | + | </center> |
| - | <br>
| + | |
| - | <br>
| + | |
| | <br> | | <br> |
| | <br> | | <br> |
| | <b>Exp 3</b> | | <b>Exp 3</b> |
| | + | <center> |
| | <table border=1> | | <table border=1> |
| | <tr align=center> | | <tr align=center> |
| Line 135: |
Line 132: |
| | </tr> | | </tr> |
| | </table> | | </table> |
| - | <br>
| |
| | <br> | | <br> |
| | [[Image:KAIST-exp3.png|260px]] | | [[Image:KAIST-exp3.png|260px]] |
| - | <br> | + | </center> |
| - | <br>
| + | |
| - | <br>
| + | |
| | <br> | | <br> |
| | <br> | | <br> |
| | <b>Exp 4</b> | | <b>Exp 4</b> |
| | + | <center> |
| | <table border=1> | | <table border=1> |
| | <tr align=center> | | <tr align=center> |
| Line 169: |
Line 164: |
| | </tr> | | </tr> |
| | </table> | | </table> |
| - | <br>
| |
| | <br> | | <br> |
| | [[Image:KAIST-exp4.png|300px]] | | [[Image:KAIST-exp4.png|300px]] |
| - | <br> | + | </center> |
| - | <br>
| + | |
| - | <br>
| + | |
| - | <br>
| + | |
| - | <br>
| + | |
| | <br> | | <br> |
| | <br> | | <br> |
Revision as of 06:56, 20 October 2010
|
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 | V_STAT1 | 68*C | - | - |
| b | B_FGFR1 | 68*C | 10% | 1398bp |
| c | VB_FGFR1 | 68*C | - | - |
Exp 8
| 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 9
| 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 |
| 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 |
9/17 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 | Selection marker(41) | 50*C | 0% | -- |
| b | Selection marker(41) | 52*C | 0% | -- |
| c | Selection marker(41) | 54*C | 0% | -- |
| d | Selection marker(41) | 56*C | 0% | -- |
| e | Selection marker(42) | 50*C | 0% | -- |
| f | Selection marker(42) | 52*C | 0% | -- |
| g | Selection marker(42) | 54*C | 0% | -- |
| h | Selection marker(42) | 56*C | 0% | -- |
| i | F_Fusion antibody receptor | 50*C | 50% | 1107bp |
| j | F_Fusion antibody receptor | 52*C | 80% | 1107bp |
| k | B_Fusion antibody receptor | 54*C | 100% | 1398bp |
| l | B_Fusion antibody receptor | 56*C | 100% | 1398bp |
Exp 2
| Segment | Annealing Temperature | Result |
| a | HA1 | 50*C | 100% | 87bp |
| b | HA1 | 52*C | 100% | 87bp |
| c | HA1 | 54*C | 100% | 87bp |
| d | HA1 | 56*C | 100% | 87bp |
| e | HA2 | 50*C | 100% | 87bp |
| f | HA2 | 52*C | 100% | 87bp |
| g | HA2 | 54*C | 100% | 87bp |
| h | HA2 | 56*C | 100% | 87bp |
| i | Proximal promoter | 50*C | 100% | 120bp |
| j | Proximal promoter | 52*C | 100% | 120bp |
| k | Proximal promoter | 54*C | 100% | 120bp |
| l | Proximal promoter | 56*C | 100% | 120bp |
| m | 5' UTR | 54*C | 100% | 500bp |
| n | 5' UTR | 54*C | 100% | 500bp |
| o | 3' UTR | 54*C | 100% | 444bp |
| p | 3' UTR | 54*C | 100% | 444bp |
9/18 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 | HA2+3'UTR | 58*C | 100% | 531bp |
| b | F_Fusion antibody receptor | 58*C | 100% | 1107bp |
| c | F_Fusion antibody receptor | 58*C | 100% | 1107bp |
| d | B_Fusion antibody receptor | 58*C | 100% | 1398bp |
| e | B_Fusion antibody receptor | 58*C | 100% | 1398bp |
360px
9/19 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 | Fusion antibody receptor | 58*C | 50% | 2493bp |
360px
9/21 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 | F_Fusion antibody receptor | 58*C | 100% | 1107bp |
| b | B_Fusion antibody receptor | 58*C | 100% | 1398bp |
| c | Core promoter | 58*C | 0% | -- |
| d | Selection marker(42) | 58*C | 0% | -- |
360px
Exp 2
| Segment | Annealing Temperature | Result |
| a | Selection marker(41) | 58*C | 0% | -- |
| b | Selection marker(42) | 58*C | 0% | -- |
360px
10/5 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 | Ig-like | 58*C | 100% | 339bp |
| b | Sig + Ig-like | 58*C | 100% | 426bp |
| c | Antibody | 58*C | 90% | 669bp |
| d | F_FGFR | 58*C | 50% | 1095bp |
| e | B_FGFR | 58*C | 100% | 1398bp |
| f | STAT1 | 58*C | 100% | 2139bp |
|
"
