Stock No: 006385
Protocol 22048: End Point Analysis Assay - Syt1<tm3Sud>
Version 3.0

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

Wt= CCGC

Mut=tCag

 

Sequence

GGCAAGACACTGGTGATGGCTGTGTATGATTTTGA(C/t)C(G/a)(C/g)TTCTCCAA

GCACGACATCATTGGAGAGTTCAAAGTTCCTATGAACACCGTGGATTTTGGCCAC

GTCACCGAGGAGTGGCG

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19749 GCT CCC TAG CAT CCT GTG AC Forward A
19750 CAA TGA TGT CGT GCT TGG AG Reverse A
19751 Fluorophore-1 GTG TAT GAT TTT GAC CGC TTC TC Quencher-1 WT Probe
19752 Fluorophore-2 TGT GTA TGA TTT TGA TCA GTT CTC C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
19749 0.40 uM
19750 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.