Stock No: 024574
Protocol 35132: End Point Analysis Assay - Gba<tm1.1Eginn>-EP
Version 1.0

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

Mut = CC

WT= TT

Sequence

gagtctggctgtgttgcccagggaagcctggggtcctgatgatttctatcttcacgttcagCAAGTTCATTCCCGAGGGATCCCAGAGAGTGGCGTTGGTGGCCAGTGAGAGCACTGAC(TT/CC)GGAAACAGTAGCACTGTTACGCCCTGACGGCTCTGCAGTTGTGGTCGTGTTAAACcGgtaagctgggtgcaagggcaggtct

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
45337 ATC CCA GAG AGT GGC GTT G Forward A
45338 Fluorophore-1 AGA GCA CTG ACT TGG AAA CAG TAG Quencher-1 WT Probe
45339 Fluorophore-2 AGC ACT GAC CCG GAA ACA GTA Quencher-2 MUT Probe
oIMR1804 ACA CGA CCA CAA CTG CAG AG Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
45337 0.40 uM
oIMR1804 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.