Stock No: 017721
Protocol 22095: End Point Analysis Assay - Uqcrfs1<tm1.1Hek> End Point
Version 3.0

Notes

Mutant =TCG/TCG
Heterozygote = CCC/TCG
Wild type = CCC/CCC

Converting to 12ul, email genotyping@jax.org with jpeg

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 =TCG/TCG
Heterozygote = CCC/TCG
Wild type = CCC/CCC

Sequence

ATTTAGATCGAGTAAAGAAACCTGAATGGGTTATTCTGATAGG

TGTCTGTACTCATCTTGGTTGTGTA(CCC/TCG)ATTGCAAAC

GCAGGAGATTTTGGTGGCTATTATTGCCCCTGCCATGGGTC

ACACTATGACGCCTCTGG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
16344 Fluorophore-1 GGT TGT GTA CCC ATT GCA AAC Quencher-1 WT Probe
16345 Fluorophore-2 GGT TGT GTA TCG ATT GCA AAC Quencher-2 MUT Probe
16346 GGA GAT TGA CCA GGA AGC TG Forward A
16347 CAT GGC AGG GGC AAT AAT AG Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
16346 0.40 uM
16347 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 40.0 -- 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.