Stock No: 002232
Protocol 32600: End Point Analysis Assay - Pah<enu2> Alt 1
Version 2.0

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

Mut = C

WT= T

CCTGGCCTTCCGAGTCTt/c

 

Sequence

CTGTTGCTGGCTTACTGTCGTCTCGAGATTTCTTGGGTGGCCTGGCCTTCCGAGTCTtCCACTGCACACAGTACATTAGGCATGGATCTAAGCCCATGTACACA

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
22621 Fluorophore-1 TCC GAG TCT TCC ACT GCA C Quencher-1 WT Probe
22622 Fluorophore-2 TCC GAG TCT CCC ACT GCA C Quencher-2 MUT Probe
39414 CTG TTG CTG GCT TAC TGT CG Forward A
39415 TGT GTA CAT GGG CTT AGA TCC Reverse A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
39414 0.40 uM
39415 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

Stock Number Strain Name
029218 B6.BTBR-Pahenu2/MalnJ
002232 BTBR-Pahenu2/J
2 strains use this protocol