Stock No: 000654
Protocol 21935: End Point Analysis Assay - Crb1<rd8>End Point
Version 3.0

Notes


Mutant = -/-  B6N (mutation fixed on this background)
Heterozygote = C/-
Wild type = C/C B6J

Use controls from DNA Resorces jr 3392

Mutation is a deleted C


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 = -/-
Heterozygote = C/-
Wild type = C/C

Mutation is a deleted C

 

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
16217 TTG CAT GGA GGA AAC TGT GA Forward A
16218 TGT ATC CAG GCT CAC ACC TG Reverse A
16219 Fluorophore-1 CAG TTC TTA TCG GTG TGC CTG Quencher-1 WT Probe
16220 Fluorophore-2 CAG TTC TTA TGG TGT GCC TG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
16217 0.40 uM
16218 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
030931 129S6.Cg-Pex1tm1.1Sjms/Mmjax
009106 B6.129-Cyp27a1tm1Elt/J
019141 B6.129S6(FVB)-S1pr1tm2.1Rlp/J
025065 B6.Cg-Pex1tm1.1Sjms/Mmjax
008451 B6.Cg-Ptprca Cx3cr1tm1Litt/LittJ
004852 B6;129-Crb1rd8/J
005304 C57BL/6NJ
000654 CBA/CaJ
003392 STOCK Crb1rd8/J
9 strains use this protocol