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Contents 1 1. Extraction of mRNA 2 2. RT-PCR 3 3.PCR 3.1 (1) Colony PCR 3.2 (2) PCR with DNA as template

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.