Stock No: 004152
Protocol 20785: End Point Analysis Assay - Ctnnb1<tm2Kem>-alternate2
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 = FAM

Wild type = HEX

Sequence

Mutant sequence:
GATGAAAGTTGTTTGTACAGAGTGTGGAGTTACTAAGTTCTTAGTATGcatgAATTCGGATCCCTCGACCGGAATTCCGATCAT
ATTCAATAACCCTTAATATAACTTCGTATAATGTATGCTATACGAAGTTATTAGGTCCCTCGAGatgCATGTGCGCGCGCGCGC
GCACACACACACAGGCTCAAAGATTTGTGTCTTTAGACATTGAATCTGTGTGTACAGTACGGTGCTAAGCATTGAAGGGAT
GGGGAGCAGCCTCATA

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
30489 Fluorophore-1 TAG TAT GCA TGT GCG CGC Quencher-1 WT Probe
30490 Fluorophore-2 ATT CCG ATC ATA TTC AAT AAC CCT T Quencher-2 MUT Probe
31042 AAA GTT GTT TGT ACA GAG TGT GGA Forward A
31043 AAG ACA CAA ATC TTT GAG CCT GTG Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
31042 0.40 uM
31043 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 40.0 -- 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.