H-ras LSL G12D PCR

Reaction Mix:

Tail DNA 2.0ul
10x buffer 2.0ul
F2 0.5ul
R1 1.0ul
SD5 0.5ul
milliQ 13ul
House Taq 1.0ul

PCR Primers:

F2 5' cca gcc atc cct cgc gtt cc 3'
R1 5’ gca ggg ctg aca gct gag agc 3’
SD5 5’ agc tag cca cca tgg ctt gag taa gtc tgc a 3’

PCR Conditions:

1. 94C 2:00
2. 94C 0:30
3. 60C 0:30
4. 72C 0:40
5. Goto 2 x34 cycles
6. 72C 5:00
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:

Wild type 391 bp

LSL 303 bp