Stock No: 009345
Protocol 12812: End Point Analysis Assay - Mstn<lean>-EP
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 =C/C
Heterozygote = T/C
Wild type = T/T

Sequence

ATGAGCCCAGGCACTGGTATTTGGCAGAGTATTGATGTGAAGACAGTGTTGCAAAA
TTGGCTCAAACAGCCTGAATCCAACTTAGGCATTGAAATCAAAGCTTTGGATGAGA
ATGGCCATGATCTTGCTGTAACCTTCCCAGGACCAGGAGAAGATGGGCTGG(t/c)AAG
TGATAACTGAAAATAACATTCTAGCAACCGTGTTACATTTTTATTCCTGACATGAATG
ATAATAGTGAAAAAGAAATACTATTTCCTATGCTCATAAGCCAGACAAAGTATCTTA
CTCCAAAGGTAGGTGTGTACCCAAAACAGTACACACCTACCTTTGGTGTGTCAATC

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
21329 ACA GCC TGA ATC CAA CTT AGG Forward
21330 AAA TGT AAC ACG GTT GCT AGA ATG Reverse
21331 Fluorophore-1 AGA TGG GCT GGT AAG TGA TAA CT Quencher-1
21332 Fluorophore-2 GAT GGG CTG GCA AGT GAT AAC Quencher-2

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 0.42
ddH2O
Primer 1 0.03
Primer 2 0.03
Primer 3 0.02
Primer 4 0.02
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 -- repeat steps 2-3 for 40 cycles
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.