Stock No: 018430
Protocol 40858: End Point Analysis Assay - Bcl2<tm1.1Sjk>
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 =GcgG
Heterozygote = (A/G)cg(T/G)
Wild type = AcgT

chr1:106712600-106712731 132bp CTTCCAGCCTGAGAGCAAC AGGGTCAGATGGACCACA

Sequence

TAAGCTGTCACAGAGGGGCTACGAGTGGGATGCTGGAGATGCGGACGCGGCGCCCCTGGGGG
CTGCCCCCACCCCTGGCATCTTCTCCTTCCAGCCTGAGAGCAACCCAATGCCCGCTGTGCACC
GGGACATGGCTGCCAGG(a/g)CG(t/g)CTCCTCTCAGGCCCCTCGTTGCCACCGCTGGGCCTGC
GCTCAGCCCTGTGCCACCTGTGGTCCATCTGACCCTCCGCCGGGCTGGGGATGACTTCTCTC
GTCGCTACCGTCGTGACTTCGCAGAGATGTCCAGTCAGCTGCACCTGA

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
56955 Fluorophore-1 CTG CCA GGA CGT CTC CTC TC Quencher-1 WT Probe
56956 Fluorophore-2 CCA GGG CGG CTC CTC T Quencher-2 MUT Probe
56957 CTT CCA GCC TGA GAG CAA C Forward A
56958 AGG GTC AGA TGG ACC ACA Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
56957 0.40 uM
56958 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.