Stock No: 010539
Protocol 23511: Standard PCR Assay - Fgf17<tm1Dor>
Version 1.2

Notes

Ran first set of samples on both the Generic Neo and Generic Cre assays and it is apparent this strain carries both cassettes. CRB 7-9-09
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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
8815 GAA GTC TCT CCA GCG ATG GGA GCC GCC CG Wild type Forward A
8816 GAC AGC AGA GAA TCA ATA GCT GCA AGC ACC Wild type Reverse A
8817 GCT ATT TTC CAT GAG TGA ACG AAC CTG GTC Mutant Forward A
8818 TTG GCT TCT CTG GGA CTC TAC CTC AGG GGC Mutant Reverse A

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
8815 0.50 uM
8816 0.50 uM
8817 0.50 uM
8818 0.50 uM
Glycerol 6.50 %
Dye 1.00 X
Kapa 2G HS taq polym 0.03 U/ul
DNA

Cycling

Step Temp °C Time Note
1 94.0 --
2 94.0 --
3 65.0 -- -0.5 C per cycle decrease
4 68.0 --
5 -- repeat steps 2-4 for 10 cycles (Touchdown)
6 94.0 --
7 60.0 --
8 72.0 --
9 -- repeat steps 6-8 for 28 cycles
10 72.0 --
11 10.0 -- hold
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.
JAX uses a 'touchdown' cycling protocol and therefore has not calculated the optimal annealing temperature for each set of primers.

Strains Using This Protocol

This is the only strain that uses this protocol.