NYMU Taipei/Lab Notes/2007 9 30
From 2007.igem.org
< NYMU Taipei/Lab Notes(Difference between revisions)
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|1 | |1 | ||
|TAT_A-INS_A | |TAT_A-INS_A | ||
| - | | | + | |209 |
|TAT_F+INS_A_R | |TAT_F+INS_A_R | ||
|TAT_A,INS_A PCR product@9/29 | |TAT_A,INS_A PCR product@9/29 | ||
| Line 16: | Line 16: | ||
|2 | |2 | ||
|TAT_B-INS_B | |TAT_B-INS_B | ||
| - | | | + | |233 |
|TAT_F+INS_B_R | |TAT_F+INS_B_R | ||
|TAT_B,INS_B PCR product@9/29 | |TAT_B,INS_B PCR product@9/29 | ||
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| - | {| 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) | ||
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Latest revision as of 13:29, 1 October 2007
PCR for TAT_A-INS_A TAT_B-INS_B
| size | Primer | Template | ||
|---|---|---|---|---|
| 1 | TAT_A-INS_A | 209 | TAT_F+INS_A_R | TAT_A,INS_A PCR product@9/29 |
| 2 | TAT_B-INS_B | 233 | 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
