Protocol 6196 - Atp11c<m2Btlr>

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

Sequence

ATTCATGGTTCTTTTCAACTTCATTA(t/a)ACCTGTCTCCATGTATGTCACAGTAGAAA

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	Primer Type
11847		GAG CTT TCT GGC ATT TTA ATT CA	Forward
11848	Fluorophore	ATG TAT TAA CCA AGG CTC CTT CAT	Reverse
11849		TGG TTC TTT TCA ACT TCA	SEQ

Reaction A

Component	Final Concentration
ddH2O
KAPA LR HotStart Buf Buffer does not contain MgCl2.	0.50
MgCl2	2.00
dNTP	0.13
KAPA LR HotStart Taq Initial Denature 3 Min This is a two enzyme system. One with proofreeading the other without. HotStart is provided by antibody.	0.01
DNA

Cycling

Step	Temp °C	Time	Note
1	94.0	--
2	94.0	--
3	60.0	--
4	65.0	--	Go to step 2-4, 50 times
6	65.0	--
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.