Stock No: 033193
Protocol 35752: End Point Analysis Assay - Adcy5<em#(R1014C)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

Wild type = C, C

Sequence

GGTGGCGCTGAAGGTGGTGACGCCCATCATCATCTCTGTCTTCGTGCTGGCTCTGTATCTGCATGCCCAGCAGGTGGAGTCCACTGCT(c/t)G(c/t)CTTGACTTCCTCTGGAAACTGCAGgtgggtggccctgtactagagttgcacagggaaagcggcaaacctcacagcacttactag

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
46941 TCG TGC TGG CTC TGT ATC TG Forward A
46942 ACC CAC CTG CAG TTT CCA G Reverse A
46943 Fluorophore-1 CCA CTG CTC GCC TTG ACT T Quencher-1 WT Probe
46944 Fluorophore-2 TCC ACT GCT TGT CTT GAC TTC C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
46941 0.40 uM
46942 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.