Genotyping Protocols Database
Protocol 31544: Probe Assay - Abca7 Founder 17-3 (KI + 33bp del) - EP
Version 1.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.

 

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

Mut= 96 bp

Wt= 129 bp

Sequence

TATCTGAAG(c/g)TacactgtgagtccatccatttactatcagtggTCATGTTGCTCA

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
37121 TGA ACT TGA CCA AGG AGC AG Forward A
37122 TTT GTC ACT AGC CCT GAG CA Reverse A
37123 Fluorophore-1 AGC TAC ACT GTG AGT CCA TCC A Quencher-1 WT Probe
37124 Fluorophore-2 TGA AGG TCA TGT TGC TCA GG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
37121 0.40 uM
37122 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.