Stock No: 008776
Protocol 23413: Standard PCR Assay - Zbtb7b<tm2Litt> STD PCR
Version 1.2

Notes

Difference between strain JR#8776 and JR# 9369 is that JR#8776 has a (GFP) attached to sequence and can be distinguished by running generic GFP assay, and JR#9369
does not.
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 = 440 bp

Heterozygote = 440 bp and 398 bp

Wild Type = 398 bp

3%

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR9667 AGA GCT GTT GCA GTC TCT AGT AGC Forward A
oIMR9668 TAC GGC CCA GTG CCT AAC AC Reverse A

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTPS-kapa 0.26 mM
oIMR9667 0.50 uM
oIMR9668 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

Stock Number Strain Name
027663 B6.129P2(Cg)-Zbtb7btm2Litt/J
009369 B6.Cg-Zbtb7btm1.1Litt/J
008776 B6;129P2-Zbtb7btm2Litt/J
3 strains use this protocol