Stock No: 004518
Protocol 5239: Sanger sequencing Assay - Agtpbp1<pcd-5J>
Version 1.0

Notes

Mut; (insert gac)
TTCCCGCACAAGGACgacGATGTCTGCTATTTC

WT;
TTCCCGCACAAGGACGATGTCTGCTATTTC

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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
11036 CGT CTG TGC ATG GAA AAG AA Forward
11037 CCT GCA GAG TCG AGT ACG TG Reverse

Reaction A

Component Final Concentration
ddH2O
High Fidelity Buffer 1.00
dNTP 0.20
MgCl2 1.50
11036 0.80
11037 0.80
Platinum Taq Pol. 0.05
DNA

Cycling

Step Temp °C Time Note
1 94.0 --
2 94.0 --
3 62.0 --
4 72.0 --
5 -- Go to step 2-4, 35 times
6 70.0 --
7 10.0 -- hold
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.