Stock No: 000219
Protocol 2097: End Point Analysis Assay - Slc30a4<lm>
Version 2.1

Notes

End point analysis is done using Amplifluor SNPS Genotyping System from Millipore. End point analysis protocols are run on an ABI 7000, 7500, or the Roche LightCycler 480
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 = red
Wild type = green
Followed by End Point Analysis on ABI 7000.

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR3450 TCC TTC ACA GTG TCA CTC AGA T Common
oIMR4229 GAA GGT CGG AGT CAA CGG ATT GTG TGC TTA TAG CTG CAT ACA TCA TAC
oIMR4230 GAA GGT GAC CAA GTT CAT GCT GTG TGC TTA TAG CTG CAT ACA TCA TAT
oIMR4231 GCC TGG CTG TCA ACT AGC TTT T Common

Reaction A

Component Final Concentration
ddH2O
Ampliflour SNP FAM 1.00 X
SNP JOE 1.00 X
Primer Mix 1.00 X
Rxn Mix S+ 1.00 X
dNTP 0.20 mM
Titanium Taq Pol. 0.05 U/ul
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 94.0 --
3 55.0 --
4 72.0 -- repeat steps 2-4 for 35 cycles
5 72.0 --
6 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.