Stock No: 010912
Protocol 37247: Probe Assay - Wt1<tm2(cre/ERT2)Wtp> Probe
Version 4.0

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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
14926 AGG ATG TTC CCC AAT GCG Wild type Forward A
14927 CCG GAA TCC TTA CCT TGG TTG Wild type Reverse A
14928 Fluorophore-1 CCT ACC TGC CCA GCT GCC T Quencher-1 WT Probe
14929 AGG AGC GGA GAA CCG TC Mutant Forward A
14930 TCT TGC GAA CCT CAT CAC TC Mutant Reverse A
14931 Fluorophore-2 CTG ACC GTA CAC CAA AAT TTG CCT GC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
14926 0.40 uM
14927 0.40 uM
14929 0.40 uM
14930 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.