Stock No: 001743
Protocol 21963: End Point Analysis Assay - Ggt1<dwg>
Version 3.0

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

WT = GCCTCGgtgatgcgaccctGTACG

Mut = GCCTCGGTACG (13 bp delection)

 

Sequence

CTACCGTGCTGAGCTGATTGAGCATCCGATGAGCATCGGCCTCGgtgatgcgaccctGTACGTGCCCAGTGCCCCACTCAGCGGGCCCGTGCTGATTCTCATCTTGAACATCCTCAAAGgtgagatgtctctctctagccctgctgaggggcagggacagagacccagtggaatgtccag

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
17546 CTA CCG TGC TGA GCT GAT TG Forward A
17547 CTG GAC ATT CCA CTG GGT CT Reverse A
17548 Fluorophore-1 CGG TGA TGC GAC CCT GT Quencher-1 WT Probe
17549 Fluorophore-2 CGG CCT CGG TAC GTG C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
17546 0.40 uM
17547 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.