Stock No: 000060
Protocol 22000: End Point Analysis Assay - Mc1r<e>
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 = -/-
Heterozygote = C/-
Wild type = C/C

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
18816 CCA GCA CCC TCT TTA TCA CC Forward A
18817 AGA AAG TGA CGA GGC AGA GC Reverse A
18818 Fluorophore-1 TAC TAC AAG CAC ACA GCC GTT CT Quencher-1 WT Probe
18820 Fluorophore-2 CTA CTA CAA GCA ACA GCC GTT CTG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
18816 0.40 uM
18817 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 4.0 -- 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.