Stock No: 032805
Protocol 34317: End Point Analysis Assay - Pacs1<em2(ins1)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 = T/T
Heterozygote = -/T
Wild type = -/- 

>chr19:5159969-5160053 85bp TTATGTTGCAAAGGAGGAAGC TCTGCCATGTTGATGAGTCC

Sequence

WT sequence with [insertion]:

CTTAAGCGAGATGCCAACAAACTGCAGATTATGTTGCAAAGGAGGAAGCGGTACAAGAACCGAACT[t]ATCTTGGGCTATAAAACCTTAGCTGTGGGACTCATCAACATGGCAGAGgtgagca

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
43301 TTA TGT TGC AAA GGA GGA AGC Forward A
43302 TCT GCC ATG TTG ATG AGT CC Reverse A
43303 Fluorophore-1 AGA ACC GAA CTA TCT TGG GCT A Quencher-1 WT Probe
43304 Fluorophore-2 AGA ACC GAA CTT ATC TTG GGC TA Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
43301 0.40 uM
43302 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.