The Jacks Lab

protocols

p107 PCR

Reaction Mix:

Tail DNA	2.0ul
10x buffer	2.0ul
p107-NEO (20uM)	0.2ul
p107-WT (20uM)	0.2ul
p107-COM (20uM)	0.4ul
milliQ	14.4ul
Taq (1/25)	1.0ul

PCR Primers:

p107-NEO	5' ACG AGA CTA GTG AGA CGT GC 3'
p107-WT	5’ TGT CCT GAG CAT GAA CAG AC 3’
p107-COM	5’ TCG CTG GCA GTC TGA GTC AG 3’

PCR Conditions:

1.	94C	3:00
2.	94C	0:30
3.	58C	1:00
4.	72C	0:45
5.	Goto 2 x34 cycles
6.	72C	5:00
7.	4C	0:00
END

10X PCR Buffer:

500 mM KCl

100mM Tris pH 8.3

15 mM MgCl₂

1 mg/ml BSA

2 mM dNTPs

dH₂0

(store at -20C)

Expected Bands:

Wt= ~280 bp

Mut= ~360 bp