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From 2012.igem.org

IrrE PCR

Gene to be amplified	Primers
irrE	Forward primer: STF1: 5'-atggggccaaaagctaaagctgaagcc-3' Reverse primer: ST2R: 5'-tcactgtgcagcgtcctgcg-3'
irrE	Forward primer: STF3 5'-gtttcttcgaattcgcggccgcttctagagatggggccaaaagctaaagctgaagcc-3' Reverse primer: ST4R 5'-gtttcttcctgcagcggccgctactagtatcactgtgcagcgtcctgcg-3'

All primers were prepared to a concentration of 1pmol.uL, from 100pmol.uL stocks. Primers ordered from MWG operon.

Component	20 µl reaction	50 µl reaction	Final concentration
Nuclease-free water	to 20 µl	to 50 µl
5X Phusion HF buffer	4 µl	10 µl	1X
10 mM dNTPs	0.4 µl	1 µl	200 µM
10 µM forward primer	1 µl	2.5 µl	0.5 µM
10 µM reverse primer	1 µl	2.5 µl	0.5 µM
Template DNA	variable	variable	< 250 ng
DMSO (optional)	(0.6 µl)	(1.5 µl)	3%
Phusion DNA polymerase	0.2 µl	0.5 µl	1.0 units/50 µl PCR