Stock No: 037610
Protocol 43838: End Point Analysis Assay - Polg<em5/6(F290F,R292C)Lutzy>
Version 1.0

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

>chr7:79460564-79460674 111bp CGGATGCGTTTTCTCGATAC CTGCTGGGTCTTGTGCTTTC

Mutant = T, T

Wild type = C, C

Sequence

SEQ

gctggcccctgtgttgcagccttccctcccctgcccttccttcagGACTCCCGGATGCGTTTTCTCGATACTATGAGCATGCACATGGCCATCTCGGGGCTGAGCAGCTT(c/t)CAG(c/t)GCAGCCTGtggatgGGAGCCAAGCAGGGAAAGCACAAGACCCAGCAGTCCACAAAGCGAGGGCAGAAGTCCCCGAGGAAAGCCAATGGTCCAGCGgtgagtgt

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
64093 CGG ATG CGT TTT CTC GAT AC Forward A
64094 CTG CTG GGT CTT GTG CTT TC Reverse A
64095 Fluorophore-1 AGC TTC CAG CGC AGC Quencher-1 WT Probe
64096 Fluorophore-2 CAG CTT TCA GTG CAG CCT Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
64093 0.40 uM
64094 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.