Stock No: 004605
Protocol 20690: End Point Analysis Assay - Itgb1<tm1Efu>-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

Mutant=  178 bp

Wild Type = 89 bp

Fam=Mut

Hex=Wt

Sequence

SEQ

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
30544 Fluorophore-1 CTC GAG GGA CCT AAT AAC TTC GT Quencher-1 MUT Probe
30888 AAA CAT TTA ATT TGC GGC TCA Forward A
30889 CCA GCT CCA TGT CTG TCT GT Reverse A
30890 Fluorophore-2 TGT CAG TCC ATG GCC GTC Quencher-2 WT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
30888 0.40 uM
30889 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

Stock Number Strain Name
034193 B6.129-Itgb1tm1Efu/RthsnJ
004605 B6;129-Itgb1tm1Efu/J
2 strains use this protocol