The Jacks Lab

protocols

p63 PCR

Reaction Mix:

Tail DNA	1.0ul
10x buffer	2.0ul
Primer 1 (20uM)	0.4ul
Primer 2 (20uM)	0.4ul
Primer 3 (20uM)	0.4ul
milliQ	14.8ul
Taq (1/25)	1.0ul

PCR Primers:

Primer 1	5' TTC TCA GAT GGT ACC GCT CC 3'
Primer 2	5' GGT GCT TTG AGG CCC GGA TC 3'
Primer 3	5' GAA AGC GAA GGA GCA AAG CTG 3'

PCR Conditions:

1.	94C	3:00
2.	94C	1:00
3.	60C	1:30
4.	72C	2:00
5.	Goto 2 x30 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= ~550 bp

Mut= ~450 bp