Today's date:	9/19/05
PCR name:	B
Purpose:	Distinguishes between wt (+) allele and Irs1 knockin mutant alleles (ki) that no longer have neo cassette at 3' end, but do have a single loxP site and other extra sequence there (66 bp total). Examples include CK, FCT, S7A, JBM segregating with + allele.
Samples:
1
2
3
4
5
		# of rxns
Reactions (ul):	X	33			Profile name:		IRS1KIGT
DNA	1				step	time (min:sec)	temp (C)
H20	13.5		445.5		1	15:00	95
10X buffer	2		66		2	:30	94
4mM dNTPs	1.5		49.5		3	:20	62
10pmol/ul primers					4	:45	72
cdsC2	1		33		go to	step 2	34 times
UTRrev1	1		33		5	10:00	72
					6	hold	4
	20	ul	627	ul
+ HotStar Taq	0.18	ul	5.94	ul
Primers:	Sequence: (5' to3')							Tm
cdsC2	CAGCAATGAGGGCAACTCCCCAAGACGCTCCA							71.7
UTRrev1	AGAGAGAAGCCCTTCTGTGGCTGCTCCAAACACA							67.5
Products:	from allele(s)		length (bp)	digests?		primers
+	wt (+)		423			cdsC2 + UTRrev1
ki	ki (CK, FCT, S7A, etc)		358			cdsC2 + UTRrev1
Genotype:	bands
+/+	358 only
CK/+, FCT/+, etc	423 and 358
FCT/FCT, CK/CK	423 only