NYMU Taipei/Lab Notes/2007 9 29
From 2007.igem.org
< NYMU Taipei/Lab Notes(Difference between revisions)
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<p> | <p> | ||
- | PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS | + | <h4>PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS</h4> |
{| align=left | border=1 | style="text-align:center" | {| align=left | border=1 | style="text-align:center" | ||
! | ! | ||
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|1.5uL | |1.5uL | ||
|- | |- | ||
- | | | + | |<i>Pfu</i> polymerase |
|0.5uL | |0.5uL | ||
|- | |- | ||
Line 113: | Line 113: | ||
</p> | </p> | ||
- | + | <br> | |
- | <p> | + | <p><hr> |
- | PCR for IDE | + | <h4>PCR for IDE</h4> |
{| align=left | border=1 | style="text-align:center" | {| align=left | border=1 | style="text-align:center" | ||
! | ! | ||
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|- | |- | ||
|IDE | |IDE | ||
- | | | + | |3105bp |
|IDE_F+IDE_R | |IDE_F+IDE_R | ||
|M_IDE | |M_IDE | ||
Line 146: | Line 146: | ||
|1.5uL | |1.5uL | ||
|- | |- | ||
- | | | + | |<i>Pfu</i> polymerase |
|0.5uL | |0.5uL | ||
|- | |- | ||
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|Denatuation<br>Annealing<br>Extension | |Denatuation<br>Annealing<br>Extension | ||
|95<br>55<br>72 | |95<br>55<br>72 | ||
- | |1<br>1<br>4 | + | |1<br>1<br><font color=red>4</font> |
|35 | |35 | ||
|- | |- | ||
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|} | |} | ||
</p> | </p> | ||
+ | <br><br> | ||
+ | <hr> | ||
+ | <p> | ||
+ | <h4>PCR for TAT_B</h4> | ||
+ | {| align=left | border=1 | style="text-align:center" | ||
+ | ! | ||
+ | ! | ||
+ | !size | ||
+ | !Primer | ||
+ | !Template | ||
+ | |- | ||
+ | |||
+ | |3 | ||
+ | |TAT_B | ||
+ | |137bp | ||
+ | |TAT_F+TAT_B_R | ||
+ | |E.coli K12 genome DNA | ||
+ | |- | ||
+ | |} | ||
+ | <br><br><br> | ||
+ | <hr> | ||
+ | <h4>2%Gel check for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS </h4> | ||
+ | [[Image:NYMU_Taipei_20070929_TAT_OmpRBS_check.jpg|300px]] | ||
+ | *Well 2 TAT_A | ||
+ | *Well 3 INS_A | ||
+ | *Well 4 TAT_B (seems <font color=red>failed</font>) | ||
+ | *Well 5 INS_B | ||
+ | *Well 6 TAT_D | ||
+ | *Well 7 OmpRBS | ||
+ | *Well 8 100bp ladder | ||
+ | |||
+ | <hr> | ||
+ | |||
+ | <h4>2%Gel check for IDE&TAT_B</h4> | ||
+ | [[Image:NYMU Taipei 20070929 IDE TATB check.jpg|300px]] | ||
+ | *Well 2 100bp ladder | ||
+ | *Well 3 IDE | ||
+ | *Well 4 TAT_B (re-PCR, TAT_B second trail) | ||
+ | *Well 5 1kb ladder |
Latest revision as of 09:42, 2 October 2007
Contents |
PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS
size | Primer | Template | ||
---|---|---|---|---|
1 | TAT_A | 140bp | TAT_F+TAT_A_R | E.coli K12 genome DNA |
2 | INS_A | 111bp | INS_A_F+INS_A_R | MGC clone (H_INS 1) |
3 | TAT_B | 137bp | TAT_F+TAT_B_R | E.coli K12 genome DNA |
4 | INS_B | 135bp | INS_B_F+INS_B_R | MGC clone (H_INS 1) |
5 | TAT_D | 131bp | TAT_F+TAT_D_R | E.coli K12 genome DNA |
6 | OmpRBS | 100bp | OmpRBS_F+OmpRBS_R | pOmpC |
Template | 1uL | |||
10X buffer | 5uL | |||
2.5mM dNTP | 2uL | |||
Primer_F | 1.5uL | |||
Primer_R | 1.5uL | |||
Pfu polymerase | 0.5uL | |||
ddH2O | 38.5uL | |||
Total | 50uL |
Step | Temp (°C) | Time (mins) | Cycle |
---|---|---|---|
Initial Denaturation | 95 | 10 | |
Denatuation Annealing Extension | 95 55 72 | 1 1 1 | 35 |
Final Extension | 72 | 10 | |
Soak | 4 |
PCR for IDE
size | Primer | Template | |
---|---|---|---|
IDE | 3105bp | IDE_F+IDE_R | M_IDE |
Template | 1uL | |||
10X buffer | 5uL | |||
2.5mM dNTP | 2uL | |||
Primer_F | 1.5uL | |||
Primer_R | 1.5uL | |||
Pfu polymerase | 0.5uL | |||
ddH2O | 38.5uL | |||
Total | 50uL |
Step | Temp (°C) | Time (mins) | Cycle |
---|---|---|---|
Initial Denaturation | 95 | 10 | |
Denatuation Annealing Extension | 95 55 72 | 1 1 4 | 35 |
Final Extension | 72 | 10 | |
Soak | 4 |
PCR for TAT_B
size | Primer | Template | ||
---|---|---|---|---|
3 | TAT_B | 137bp | TAT_F+TAT_B_R | E.coli K12 genome DNA |
2%Gel check for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS
- Well 2 TAT_A
- Well 3 INS_A
- Well 4 TAT_B (seems failed)
- Well 5 INS_B
- Well 6 TAT_D
- Well 7 OmpRBS
- Well 8 100bp ladder
2%Gel check for IDE&TAT_B
- Well 2 100bp ladder
- Well 3 IDE
- Well 4 TAT_B (re-PCR, TAT_B second trail)
- Well 5 1kb ladder