Master Protocol Details


Stock Number:	006613
Strain Name:	STOCK Tg(CAG-Bgeo,-Tle1,-ALPP)1Lbe/J





Allele:	Generic LacZ
Protocol Name:	Generic LacZ Melt Curve Analysis
Method:	Melt Curve Analysis
Version:	3.0
Created:	05-January -2010	Updated:	06-March -2013



Expected Results:	Transgene Tm = 87°C +/- 1.0°C Internal positive control Tm = 82.1°C+/- 1.0°C

Gel Image

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Protocol Primers

Primer	5' Label	Sequence 5' --> 3'	3' Label	Primer Type
oIMR0040	-	CGT GGC CTG ATT CAT TCC	-	Transgene Reverse
oIMR3054	-	ATC CTC TGC ATG GTC AGG TC	-	Transgene Forward
oIMR8744	-	CAA ATG TTG CTT GTC TGG TG	-	Internal Positive Control Forward
oIMR8745	-	GTC AGT CGA GTG CAC AGT TT	-	Internal Positive Control Reverse


				Number of Reactions

Reaction/Components

Reaction Component	Volume (µl)	Final Concentration	Total Volume (µl)
ddH2O	2.80	-	2.80
2 X SYBR Green Master Mix (Roche)	6.00	1.00 X	6.00
20 uM oIMR3054	0.30	0.50 uM	0.30
20 uM oIMR0040	0.30	0.50 uM	0.30
20 ul oIMR8744	0.30	0.50 ul	0.30
20 uM oIMR8745	0.30	0.50 uM	0.30
DNA	2.00	-	2.00

Cycling

Step #	Temp °C	Time	Note
1	94	5 min	-
2	94	30 sec	-
3	50	45 sec	-
4	72	30 sec	repeat steps 2-4 for 35 cycles
5	72	2 min	-
6	4	-	hold

This genotyping protocol is used for the following strains:

Stock Number	Strain Name
002621	B6;SJL-Tg(tetop-lacZ)2Mam/J
002856	FVB/N-Tg(TIE2-lacZ)182Sato/J
003299	B6;SWJ-Tg(TIMP3-lacZ)7Jeb/J
004141	B6;CBA-Tg(UAS-lacZ)65Rth/J
004623	STOCK Tg(Fos-lacZ)34Efu/J
005317	B6.Cg-Tg(BAT-lacZ)3Picc/J
005493	STOCK Tg(Tek-rtTA,TRE-lacZ)1425Tpr/J
005728	STOCK Tg(tetO-Ipf1,lacZ)958.1Macd/J
005814	B6.129P2-Grm1^tm1Dgen/J
006229	B6.Cg-Tg(DRE-lacZ)2Gswz/J
006465	B6;CBA-Tg(CAG-lacZ-WGA)330Bbm/J
006477	B6.Cg-Tg(CAG-lacZ-WGA)330Bbm/J
006578	STOCK Myoz2^tm1Eno/J
006613	STOCK Tg(CAG-Bgeo,-Tle1,-ALPP)1Lbe/J
006665	B6;129P2-Olfr151^tm13(rI7)Mom/MomJ
006672	B6;CBA-Tg(Olfr16*,taulacZ)7Mom/MomJ
006686	STOCK Olfr6^tm1Mom/MomJ
007972	B6;CBA-Tg(Olfr151-taulacZ)4Mom/MomJ
007973	B6;CBA-Tg(Olfr16-taulacZ)1Mom/MomJ
007974	B6;CBA-Tg(Olfr160-taulacZ)V4-7Mom/MomJ
007975	B6;CBA-Tg(OR8A1-taulacZ)1Mom/MomJ
007976	B6;CBA-Tg(Olfr713-taulacZ)4Mom/MomJ
008344	B6;DBA-Tg(Fos-tTA,Fos-EGFP)1Mmay Tg(tetO-lacZ,tTA)1Mmay/J
008477	STOCK Tg(RARE-Hspa1b/lacZ)12Jrt/J
008604	FVB.129(B6)-Smn1^{tm5(Smn1/SMN2)Mrph}/J
008704	B6;129-Smn1^{tm6(SMN2)Mrph}/J
009348	B6.129P2(Cg)-Hprt^{tm17(Ple48-lacZ)Ems}/Mmjax
011052	B6;129S4-Ctbp2^{Gt(ROSA61)Sor}/J
012333	B6.129P2(Cg)-Hprt^{tm45(Ple67-lacZ)Ems}/Mmjax
012347	B6.129P2(Cg)-Hprt^{tm64(Ple170-lacZ)Ems}/Mmjax
012354	STOCK Hprt^{tm66(Ple5-lacZ)Ems}/Mmjax
012441	STOCK Tg(tetO-LRRK2*G2019S)E3Cai/J
012575	B6.129P2(Cg)-Hprt^{tm39(Ple24-lacZ)Ems}/Mmjax
012583	B6.129P2(Cg)-Hprt^{tm68(Ple127-lacZ)Ems}/Mmjax
012584	STOCK Hprt^{tm69(Ple134-lacZ)Ems}/Mmjax
012656	B6.129P2(Cg)-Hprt^{tm70(Ple240-lacZ)Ems}/Mmjax
012657	B6.129P2(Cg)-Hprt^{tm71(Ple155-lacZ)Ems}/Mmjax
012659	B6.129P2(Cg)-Hprt^{tm73(Ple142-lacZ)Ems}/Mmjax
012733	B6.129P2(Cg)-Hprt^{tm53(CAG-lacZ)Ems}/Mmjax
012734	B6.129P2(Cg)-Hprt^{tm74(Ple232-lacZ)Ems}/Mmjax
012850	B6;129P2-Tardbp^{Gt(RRB030)Byg}/J
013764	STOCK Hprt^{tm57(Ple26-lacZ)Ems}/Mmjax
017524	B6;129-Tg(CMV-Bgeo,-WGA,-ALPP)1Mgmj/J
018054	B6.129P2(Cg)-Pitx2^tm1.1Dmm/J
018554	B6N(Cg)-Gpnmb^{tm1.1(KOMP)Vlcg}/J
018561	B6N(Cg)-Ahrr^{tm1b(KOMP)Wtsi}/J
018587	B6N(Cg)-Cited4^{tm1.1(KOMP)Vlcg}/J
018625	B6.FVB-Tg(Fabp4-lacZ)4Mosh/J
018627	B6;SJL-Tg(Myl1-lacZ)1Ibdml/J
019146	B6.129-Tmc1^tm1.1Ajg/J
304623	Tg(Fos-lacZ)34Efu/J

Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.