Stock No: 013758
Protocol 18776: Standard PCR Assay - Batf<tm1.1Kmm>
Version 3.0

Notes

This assay does not work well without the use of a Hotstart Taq polymerase.
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 = 470 bp
Heterozygote = 283 bp and 470 bp
Wild type = 283 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
11392 TGC TGA GCA TCT TTC CAG TCC Common
11393 AGA ATC CAG TGA CTC CCT ATC CC Wild type
11394 CTG CTC AGT CTC AGT TTT CAC Mutant

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
Primer 1 0.50 uM
Primer 2 0.50 uM
Primer 3 0.50 uM
Glycerol 6.50 %
Dye 1.00 X
Kapa 2G HS taq polym 0.03 U/ul
DNA

Cycling

Step Temp °C Time Note
1 94.0 --
2 94.0 --
3 65.0 -- -1.5 C per cycle decrease
4 68.0 --
5 -- repeat steps 2-4 for 10 cycles
6 94.0 --
7 50.0 --
8 72.0 --
9 -- repeat steps 6-8 for 28 cycles
10 72.0 --
11 10.0 -- hold
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
013757 129S-Batftm1.1Kmm/J
013758 B6.129S-Batftm1.1Kmm/J
013759 C.129S-Batftm1.1Kmm/J
3 strains use this protocol