NYMU Taipei/Lab Notes/2007 9 30
From 2007.igem.org
(Difference between revisions)
m |
|||
Line 22: | Line 22: | ||
|} | |} | ||
- | {| align=center | border=1 | style="text-align:center" | + | {|align=left| border=1 | style="text-align:center" |
+ | |||
+ | |Template1 | ||
+ | |1uL | ||
+ | |- | ||
+ | |Template2 | ||
+ | |1uL | ||
+ | |- | ||
+ | |10X buffer | ||
+ | |5uL | ||
+ | |- | ||
+ | |2.5mM dNTP | ||
+ | |2uL | ||
+ | |- | ||
+ | |Primer_F | ||
+ | |1.5uL | ||
+ | |- | ||
+ | |Primer_R | ||
+ | |1.5uL | ||
+ | |- | ||
+ | |<i>Pfu</i> polymerase | ||
+ | |0.5uL | ||
+ | |- | ||
+ | |ddH2O | ||
+ | |37.5uL | ||
+ | |- | ||
+ | |Total | ||
+ | |50uL | ||
+ | |-} | ||
+ | |||
+ | |||
+ | {| align=left | border=1 | style="text-align:center" | ||
!Step | !Step | ||
!Temp (°C) | !Temp (°C) | ||
Line 55: | Line 86: | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
<br><br> | <br><br> |
Revision as of 18:15, 30 September 2007
PCR for TAT_A-INS_A TAT_B-INS_B
size | Primer | Template | ||
---|---|---|---|---|
1 | TAT_A-INS_A | TAT_F+INS_A_R | TAT_A,INS_A PCR product@9/29 | |
2 | TAT_B-INS_B | TAT_F+INS_B_R | TAT_B,INS_B PCR product@9/29 |
Template1 | 1uL | ||
Template2 | 1uL | ||
10X buffer | 5uL | ||
2.5mM dNTP | 2uL | ||
Primer_F | 1.5uL | ||
Primer_R | 1.5uL | ||
Pfu polymerase | 0.5uL | ||
ddH2O | 37.5uL | ||
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 |
2%Gel check for TAT_A-INS_A TAT_B-INS_B
- Well 1 TAT_A-INS_A 10uL
- Well 2 TAT_B-INS_B 10uL
- Well 3 100bp ladder 5uL