Transgene "CRE" Genotyping
Reaction Mix:
Qiagen (hotstar taq)
10X PCR Buffer	2.5 µL
Q solution	5 µL
dNTP	2 µL
CRE primer mix	1 µL
Beta 5* primer mix	3 µL
Taq	0.125 µL
DNA	1.0 µL
H2O	10.375 µL
(dissolve the DNA from 1 cm of dissected tail in 500µL of TE. (Use 1µL for reactions))
PCR Primers:
CRE(F)	5'-ACTATCCAGCAACATTTGGGCCAG-3'
CRE(R)	5'-GATCCTGGCAATTTCGGCTATACG-3'
internal control
*Beta5(F1)	5'-CTACAGCATACCTGCTTCACTGTG-3'
*Beta5(R)	5'-GCCTTTCTGTGGGTTCCA-3'
Primer mix
STARTING WITH 100mM PRIMER STOCKS
8.4 µL Beta5 (F1)	25 µL CRE(F)
8.4 µL Beta5 (R)	25 µL CRE(R)
183.2 µL H2O	150 µL H2O
PCR CONDITIONS (Use ANG1-SOF):
1. 95° C,	5:00
2. 94° C,	1:00
3. 55° C,	2:00
4. 72° C,	4:00
5. Goto 2,	x34
6. 72° C,	15:00
7. END
Expected Bands
internal control B5: ~ 434 bp
transgene Mutant: ~ 282 bp