PCR Genotyping Primer Pairs

The following primer pairs will amplify sequences present as a single copy in the mouse genome with the Universal Genotyping Protocol. Note the two sets of Fabpi primers are used as internal controls. The Fabpi primers amplify sequences from the endogenous mouse intestinal fatty acid binding protein gene. Two different sized control amplimers are available so that the control band won’t interfere with the experimental band of any size.

β-galactosidase (LacZ)

5’ primer	GTT GCA GTG CAC GGC AGA TAC ACT TGC TGA
3’ primer	GCC ACT GGT GTG GGC CAT AAT TCA ATT CGC

Cre-recombinase

These primers amplify the Clontech enhanced CFP.

5’ primer	GCA TTA CCG GTC GAT GCA ACG AGT GAT GAG
3’ primer	GAG TGA ACG AAC CTG GTC GAA ATC AGT GCG

CFP

These primers amplify the Clontech enhanced CFP.

5’ primer	GCA GCA CGA CTT CTT CAA GTC CGC CAT GCC
3’ primer	GTG GCG GAT CTT GAA GTT GGC CTT GAT GCC

Diphtheria toxin

5’ primer	GTG GTC AAA GTG ACG TAT CCA GGA CTG ACG
3’ primer	AGA ACT ACA CGC GAA GCA CCA TCC CCG AAC

DsRED

5’ primer	GTA ATG CAG AAG AAG ACT ATG GGC TGG GAG
3’ primer	ATG TCC AGC TTG GAG TCC ACG TAG TAG TAG

FABPI-200

These primers are used as internal controls and are included in reactions with other primers. They amplify a sequence from the intestinal fatty acid binding protein gene. There are two sets of FABPI primers one for a 200 base pair amplimer and one for a 500 base pair amplimer.

5’ primer	TGG ACA GGA CTG GAC CTC TGC TTT CCT AGA
3’ primer	TAG AGC TTT GCC ACA TCA CAG GTC ATT CAG

FABPI-500

These primers are used as internal controls and are included in reactions with other primers. They amplify a sequence from the intestinal fatty acid binding protein gene. There are two sets of FABPI primers one for a 200 base pair amplimer and one for a 500 base pair amplimer.

5’ primer	CCT CCG GAG AGC AGC GAT TAA AAG TGT CAG
3’ primer	TAG AGC TTT GCC ACA TCA CAG GTC ATT CAG

FLP recombinase

5’ primer	GTC CAC TCC CAG GTC CAA CTG CAG CCC AAG
3’ primer	CGC TAA AGA AGT ATA TGT GCC TAC TAA CGC

GFP/BFP/YFP

These primers amplify the Clontech enhanced GFP, BFP and YFP.

5’ primer	GCA CGA CTT CTT CAA GTC CGC CAT GCC
3’ primer	GCG GAT CTT GAA GTT CAC CTT GAT GCC

Human growth hormone (complete)

These primers are near the 5’ end of the entire hGH gene.

5’ primer	CTG CAC CAG CTG GCG TTT GAC ACC TAC CAG
3’ primer	TTT CTG TTG TGT TTC CTC CCT GTT GGA GGG

Human growth hormone (transcriptional stop)

These primers amplify the hGH sequence surrounding the polyA signal, which is commonly used in transgenes as a transcriptional stop sequence.

5’ primer	ACT CCA GTG CCC ACC AGC CTT GTC CTA ATA
3’ primer	ATT GCA GTG AGC CAA GAT TGT GCC ACT GCA

Luciferase (click-beetle)

5’ primer	GTC GAT GTG GTC GGC GAT GAA TCT TTG AGC
3’ primer	TAG CTC TCG TTG ACT GGA GCC ACG ATC ATA

Luciferase (firefly)

5’ primer	AGA CGC CAA AAA CAT AAA GAA AGG CCC GGC
3’ primer	TAT AAA TGT CGT TCG CGG GCG CAA CTG CAA

Neomycin phosphotransferase

5’ primer	TGC TCC TGC CGA GAA AGT ATC CAT CAT GGC
3’ primer	CGC CAA GCT CTT CAG CAA TAT CAC GGG TAG

SRY (male-specific)

SRY F	TTG TCT AGA GAG CAT GGA GGG CCA TGT CAA
SRY R	CCA CTC CTC TGT GAC ACT TTA GCC CTC CGA

tTA (tet-on)

5’ primer	AGC TTG GTG TAG AGC AGC CTA CAC TGT ATT
3’ primer	GCT CCA TTG CGA TGA CTT AGT AAA GCA CAT