Stock No: 033998
Protocol 36386: End Point Analysis Assay - Dhps<em5(del7)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

Mutant = -------

Wild type = GACAATT 
>chr8:85073486+85073572 87bp ATGGCTCCCATTTCCAGGAT GTCCAGAATGGGCATGAGC

Sequence

WT sequence with deletion in lowercase:
ggctttgagtagtggtcggggcggggaagctgggcagaggatccccgtgacccatgcccgcatggctcccatttccagGATTGGAAACCTGCTGGTGCCGAATgacaattACTGCAAGTTTGAGGACTGGCTCATGCCCATTCTGGACCAGATGGTGCTGGAGCAGAACACAGAGgtggggctggaagggctcggggacctgcatagtaagag

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
48604 ATG GCT CCC ATT TCC AGG AT Forward A
48608 GTC CAG AAT GGG CAT GAG C Reverse A
48609 Fluorophore-1 TGC CGA ATG ACA ATT ACT GC Quencher-1 WT Probe
48610 Fluorophore-2 TGG TGC CGA ATA CTG CAA GT Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
48604 0.40 uM
48608 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.