Stock No: 033187
Protocol 35907: End Point Analysis Assay - Adcy5<em#(M1030K)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 = A, A

Wild type = G, T 

>chr16:35298411+35298488 78bp TCACTGGGCCTTCTGTCC AGCAGGCGCCGATTGTAG

Sequence

 gtttcaggggcattttggtctcctctcactgggccttctgtccccagGCCACAGAAGAGAA(g/a)GAGGAGA(t/a)GGAGGAGCTGCAGGCCTACAATCGGCGCCTGCTGCACAACATTCTGCCCAAGGACGTGGCTGCCCACTTCCTGGCCCGGGAACGGCGCAATGACGAGCTGTACTACCAATCGTG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
47290 TCA CTG GGC CTT CTG TCC Forward A
47291 AGC AGG CGC CGA TTG TAG Reverse A
47292 Fluorophore-1 AGG AGG AGA TGG AGG AGC TG Quencher-1 WT Probe
47293 Fluorophore-2 AAA GAG GAG AAG GAG GAG CTG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
47290 0.40 uM
47291 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.