Strain Protocols

Strain Information
Strain Code: 01XJ1
Strain Name: B6.129S7(Cg)-Mdm2tm1.1Mep/Nci
Gene Symbol: Mdm2
Protocol Information
Allele: Mdm2<tm2Mep>
Protocol Number: 1
Introductory Comment: This strain is commonly called mdm2rec where the exons 7-9 have been removed by LoxP recombination. The mice are maintained as heterozygotes since homozygous is embryonic lethal.

Primers

M030 : 5'-cTG TGT GAG cTG AGG GAG ATG TG-3'
M031 : 5'-ccT GGA TTT AAT cTG cAG cAc Tc-3'
M032 : 5'-cTc cAG cAT GcA AAT GAc TcT TG-3'

Product Sizes

Primer Combination Product Size
M030 / M031 310bp    Wildtype
M032 / M031 240bp    mdm2rec

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
DMSO 1%
100 µMM030 0.3 µM
100 µMM031 0.3 µM
100 µMM032 0.3 µM
5U/┬Ál Taq Pol 0.025 U/µl
Use 50 - 100 ng of DNA per reaction

Cycling Conditions

(Time in minutes:seconds)
Temperature Time Label
94ºC 3:00
0ºC
94ºC 1:00
60ºC 2:00 35 Cycles
72ºC 1:00
72ºC 3:00
4ºC HOLD

Disclaimer: PCR conditions were optimized using QIAGEN-purified mouse tail DNA and a PTC-200 thermocycler (MJ Research). Optimal conditions may vary with DNA quality or equipment used.