Protocol 41853: Probe Assay - Adora2a<tm1Jfc>-alt1
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

Mutant=  79 bp

Wild Type = 83 bp

>chr10:75326318+75326400 83bp TCGACAGATACATCGCCATC TTTGTGCCCACAGATCTAGC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
59285 TCG ACA GAT ACA TCG CCA TC Common A
59286 TTT GTG CCC ACA GAT CTA GC Wild type Reverse A
59287 Fluorophore-1 TCC ACT CCG GTG AGC CAG Quencher-1 WT Probe
59288 Fluorophore-2 TCT ACC GGG TAG GGG AGG C Quencher-2 MUT Probe
oIMR5316 CTA AAG CGC ATG CTC CAG AC Mutant Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
59285 0.40 uM
59286 0.40 uM
oIMR5316 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

Stock Number Strain Name
010685 C;129S-Adora2atm1Jfc/J
034539 STOCK Adora1tm1Jgsc Adora3tm1Jbsn Adora2atm1Jfc Adora2btm1Till/ReiMmjax
2 strains use this protocol