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Team:Nanjing-China/Notebook
From 2013.igem.org
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Contents |
1. Extraction of mRNA
- Kit
2. RT-PCR
- Kit
3.PCR
(1) Colony PCR
2×PrimeStar 12.5μl Forward primer 1~2.5μl Reverse primer 1~2.5μl Bacteria solution 1~2μl dd water up to 25μl
Step 1 94℃ Step 2 94℃ Step 3 Annealing temperature Step 4 72℃ Step 5 goto step2, 25~35 cycles Step 6 72℃ Pause at 4℃
(2) PCR with DNA as template
2×PrimeStar 12.5μl Forward primer 1μl Reverse primer 1μl Template DNA 0.5μl dd water up to 25μl
Step 1 94℃ Step 2 94℃ Step 3 Annealing temperature Step 4 72℃ Step 5 goto step2, 25~35 cycles Step 6 72℃ Pause at 4℃
- Gel running and purify the product.
