The Jacks Lab

protocols

p21 PCR

Reaction Mix:

Tail DNA	2.0ul
10x buffer	2.0ul
+116F (20umol)	1.0ul
135 (20umol)	1.0ul
Neo 19+ (20umol)	1.0ul
milliQ	12.0ul
Taq (1/25)	1.0ul

PCR Primers:

+116F	5' AAG CCT TGA TTC TGA TGT GGG C 3'
135	5’ TGA CGA AGT CAA AGT TCC ACC G 3’
Neo 19+	5’ GCT ATC AGG ACA TAG CGT TGG C 3’

PCR Conditions:

1.	94C	4:00
2.	94C	1:00
3.	60C	1:00
4.	72C	2:00
5.	Goto 2 x35 cycles
6.	72C	7:00
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= ~900 bp

Mut= ~759 bp