Stock No: 002650
Protocol 28088: Standard PCR Assay - Il6<tm1Kopf> MCA
Version 2.2

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

Mut tm 89°
Wt  tm 85°


HET = 380 bp and 174 bp
MUT = 380 bp
WT = 174 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR0212 TTC CAT CCA GTT GCC TTC TTG G Common A
oIMR0213 TTC TCA TTT CCA CGA TTT CCC AG Wild type Reverse A
oIMR0214 CCG GAG AAC CTG CGT GCA ATC C Mutant Reverse A Neo R

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
oIMR0212 0.50 uM
oIMR0213 0.50 uM
oIMR0214 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
002650 B6.129S2-Il6tm1Kopf/J
002254 B6;129S2-Il6tm1Kopf/J
021873 BXSB.129S2(B6)-Il6tm1Kopf/DcrJ
007078 CByJ.129S2(B6)-Il6tm1Kopf/J
029264 NOD.129S2(B6)-Il6tm1Kopf/YgchJ
5 strains use this protocol