The Jacks Lab

protocols

Nf2 PCR

Reaction Mix (store at -20C):

100 pmol stock NF2WT	20 ul
100 pmol stock NF2X2	20 ul
100 pmol stock neo18	20 ul
10 X buffer (see below)	500 ul
milliQ H2O	4430 ul

For each reaction

Tail DNA	2 ul
RX Mix	17 ul
Taq (1/25)	1.0ul

PCR Primers:

Nf2WT	5' GTC TGG GAA GTC TGT GGA GG 3'
Nf2X2	5’ GGG GCT TCG GGA AAC CTG G 3’
Neo18	5’ CTA TCA GGA CAT AGC GTT GG 3’

PCR Conditions:

1.	94C	1:00
2.	94C	0:30
3.	61C	1:45
4.	72C	1:45
5.	Go to 2 x29 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 = 306 bp

Mut = 574 bp