Stock No: 026012
Protocol 24521: Standard PCR Assay - Apoe<tm1Unc>
Version 4.2

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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR0180 GCC TAG CCG AGG GAG AGC CG Common A
oIMR0181 TGT GAC TTG GGA GCT CTG CAG C Wild type Reverse A
oIMR0182 GCC GCC CCG ACT GCA TCT 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
oIMR0180 0.50 uM
oIMR0181 0.50 uM
oIMR0182 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
014556 129S6/SvEv-Apoetm4Mae/J
029769 B6.129-Apoetm1Unc Apptm1Dbo/Mmjax
002246 B6.129-Apoetm1Unc Ldlrtm1Her/J
002052 B6.129P2-Apoetm1Unc/J
026016 B6.129P2-Uts2tm1Ags Apoetm1Unc/J
004631 B6.Cg-Apoetm1Unc Cdh18Tg(GFAP-APOE_i4)1Hol/J
007224 B6.Cg-Apoetm1Unc (D12Mit182-D12Mit2)/Pgn
027052 B6.Cg-Apoetm1Unc Tg(MSR1-TXN*)23Arem/Mmjax
008525 B6.Cg-Tg(Cyp21a1-Apoe)619Fet/J
004632 B6.Cg-Tg(GFAP-APOE_i2)14Hol Apoetm1Unc/J
004633 B6.Cg-Tg(GFAP-APOE_i3)37Hol Apoetm1Unc/J
002878 B6;129-Apobtm1Sgy Apoetm1Unc/J
002879 B6;129-Apobtm2Sgy Apoetm1Unc/J
002245 B6;129-Apoetm1Unc Ldlrtm1Her/J
004362 B6;129-Scarb1tm1Kri Apoetm1Unc/J
026012 B6;129P2-Uts2rtm1Djbe/J
030895 B6NCrl.129(B6)-Apoetm1Unc Ccr1tm1Gao Ccr5tm1Blck/BlckJ
030892 B6NCrl.129(B6J)-Apoetm1Unc Ccr1tm1Gao/BlckJ
030896 B6NCrl.129P2(B6)-Apoetm1Unc Ccr2tm1.1Blck Ccr5tm3.1Blck/BlckJ
030893 B6NCrl.129P2(B6)-Apoetm1Unc Ccr2tm1.1Blck/BlckJ
030894 B6NCrl.129P2(B6)-Apoetm1Unc Ccr5tm1Blck/BlckJ
030891 B6NCrl.129P2(Cg)-Apoetm1Unc/BlckJ
007067 D2.129P2(B6)-Apoetm1Unc/J
007069 STOCK Apoetm1Unc/J
24 strains use this protocol