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