Stock No: 021984
Protocol 22066: End Point Analysis Assay - Trp53<tm3Att>
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

Mutant = CAGTCT
Wild type = TTTTTT

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
17343 CAT CAC CTC ACT GCA TGG AC Forward A
17344 GAC ACT CGG AGG GCT TCA CT Reverse A
17347 Fluorophore-1 TGA GGA GTT TTT TGA AGG CC Quencher-1 WT Probe
17348 Fluorophore-2 TGA GGA GCA GTC TGA AGG CC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
17343 0.40 uM
17344 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
018431 129-Trp53tm3Att/J
021984 129-Trp53tm4Att/J
2 strains use this protocol