Stock No: 017688
Protocol 32902: Probe Assay - Shank3<tm2Gfng> Probe
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

>chr15:89531337+89531455 119bp CCGCAGAAACCAGAGAGGAC TGCTACTGTCCCTGCGTGT

Mutant=  110 bp

Wild Type = 119 bp

Sequence

Wt Sequence: ccgcagAAACCAGAGAGGACCGGACGAAGCGTCTCTTCCGCCACTACacTGTGGGTTCCTATGACAGCCTCACTTCACACAGgtactggcaggggtgtggacacgcagggacagtagca

 

Mutant Sequence: CCGCAGAAACCAGAGAGGACCGGACGAAGCGTCTCTTCCGCCACTACagGCGCGCCTAGTCGACATAACTTCGTATAGCATACATTATACGAAGTTATAGCGCGAAGTTC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19043 GAA CTT CGC GCT ATA ACT TCG Mutant Reverse A
40240 CCG CAG AAA CCA GAG AGG AC Common A
40241 TGC TAC TGT CCC TGC GTG T Wild type Reverse A
40242 Fluorophore-1 CGC GCC TAG TCG ACA TAA CT Quencher-1 MUT Probe
40243 Fluorophore-2 TGG GTT CCT ATG ACA GCC TC Quencher-2 WT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
19043 0.40 uM
40240 0.40 uM
40241 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.