Tempel Lab Protocols

FGF8 FLOX GENOTYPE

You will Need

Mouse Tail Digests

2 Primers:
EM40
EM41

PCR Reagents:
Sterile water
10x buffer w/MgCl2
10mM Nucleotide mix
FastStart taq
Primer Sequences

EM40 CTG CAG AAC GCC AAG TAC G
EM41 AGC TCC CGC TGG ATT CCT C

Procedure

PCR Rxn
Sterile Water

17.8 ul
10x PCR buffer w/MgCl2 2.5ul
10mM Nucleotide mix 0.5 ul
EM40 primer (20 uM) 1 ul
EM41 primer (20 uM) 1 ul
FastStart Taq 0.2ul
Tail Lysate 2 ul
Total Volume 25 ul
PCR Protocol

95 deg C 4:30
95 deg C
 0:30
54 deg C
 0:45 x30
72 deg C
 0:45
72 deg C 7:00
4 deg C Hold

Gel
2% Agarose + 3 ul EtBr (10mg/ml)

Bands
WildType: 300 bp
Het: 300bp & 350bp
Mutant: 350 bp