Nf1 PCR

Reaction Mix:

Tail DNA 2.0ul
10x buffer 2.0ul
NII (20uM) 0.2ul
NIII (20uM) 0.2ul
3'5' (20uM) 0.4ul
milliQ 14.2ul
Taq (1/25) 1.0ul

PCR Primers:

NII 5' GGT ATT GAA TTG AAG CAC 3'
NIII 5’ ATT CGC CAA TGA CAA GAC 3’
3'5' 5’ TTC AAT ACC TGC CCA AGG 3’

PCR Conditions:

1. 94C 3:00
2. 94C 1:00
3. 52C 1:00
4. 72C 2:00
5. Goto 2 x33 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= ~230 bp
Mut= ~350 bp