Team:University College London/Week12Joanne2

From 2012.igem.org

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'


Component 20µl Reaction 50µl Reaction Final Concentration
Nuclease-free water to 20µl to 50µl
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
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