Stock No: 022018
Protocol 21925: End Point Analysis Assay - B3gnt1<m1Ddg>End Point
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 = C/C
Heterozygote =T/C
Wild type = T/T

Sequence

GGAGGCGCAGCTGGCCACGGTGCTGGCCTACGCGCTGAGTAGCCACTGCCCCGAGATGCG

CGCCAGGGTCGCCATGCACCTCGTGTGCCCCTCGCGCTATGAGGCTGCTGTGCCCGACCC

CCGAGAACCTGGAGAGTTTGCCCTGCTGCGGTCCTGCCAAGAGGTCTTTGACAAGCTAGC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
16519 TGC CAG TGT GGA CAA CCT AC Forward A
16520 CAA AGA CCT CTT GGC AGG AC Reverse A
16521 Fluorophore-1 GGT CGC CAT GCA CCT CG Quencher-1 WT Probe
16522 Fluorophore-2 GGT CGC CAC GCA CCT CG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
16519 0.40 uM
16520 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.