Stock No: 029910
Protocol 31810: Probe Assay - Plcg2<em2Adiuj> - EP
Version 1.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.

 

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

Mut= 95 bp

Wt= 108 bp

Sequence

 

GTGGCTCAGGACACTCCCCCCacggagctgcattTTGGGGAGAAATGGTTCCACAAGAAG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
29569 AAT AAG CCC TTT CGG ATG CT Forward A
37723 CTT CTC CGC ACT GGT CCT AC Reverse A
37724 Fluorophore-1 CAC GGA GCT GCA TTT TGG Quencher-1 WT Probe
37725 Fluorophore-2 CCT TGG GGA GAA ATG GTT C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
29569 0.40 uM
37723 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 4.0 -- 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.