The Jacks Lab

protocols

p130 PCR

Reaction Mix:

Tail DNA	2.0ul
10x buffer	2.0ul
RB-PGK3' (20uM)	0.2ul
122-WT (20uM)	0.2ul
p130C1 (20uM)	0.4ul
milliQ	14.4ul
Taq (1/25)	1.0ul

PCR Primers:

RB-PGK3'	5' GAA GAA CGA GAT CAG CAG C 3'
122-WT	5’ TAC ATA GTT TCC TTC AGC GG 3’
p130C1	5’ ACG GAT GTC AGT GTC ACG 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= ~220 bp

Mut= ~300 bp