Stock No: 022465
Protocol 31829: End Point Analysis Assay - Pgr<tm2Omc>-Alt 2
Version 4.0

Notes

The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX). To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility. Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.

Expected Results

Mutant = C/C
Heterozygote = A/C
Wild type =A/A

Sequence

CTCCTCCCAGGAGACAGGGGAGGAGAAAAGGGGAGCTT

GGGTCGTC(a/c)tgactgagct(g/c)caggcaaaGGATCCGCAG

GTTCTCCACACGTCTGGCGCTTCGCCCTCCCCCCCAC

ACATCGG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18343 AGA CAG GGG AGG AGA AAA GG Forward A
18344 GCG AGA CTA CAG ACG ACA CG Reverse A
18345 Fluorophore-1 CGT CAT GAC TGA GCT GCA G Quencher-1 WT Probe
18346 Fluorophore-2 CGT CCT GAC TGA GCT CCA G Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
18343 0.40 uM
18344 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 --
4 -- repeat steps 2-3 for 40 cycles
5 40.0 -- Forever
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

This is the only strain that uses this protocol.