Strain Protocols

Strain Information
Strain Code: 01XMD
Strain Name: B6;129S-Mdm4GT(VICTR20)7Lex Mdm2tm1BayTrp53tm1Brd/Nci
Gene Symbol: Trp53
Protocol Information
Allele: Trp53<tm1Brd>/Nci
Protocol Number: 1
Introductory Comment: P53 genotyping protocol

Primers

Y691 : 5'-gtg ttt cat tag ttc ccc acc ttg ac-3'
Y692 : 5'-atg gga ggc tgc cag tcc taa ccc-3'
Y695 : 5'-gtg gga ggg aca aaa gtt cga ggc c-3'
Y696 : 5'-ttt acg gag ccc tgg cgc tcg atg t -3'

Product Sizes

Primer Combination Product Size
Y691 / Y692 320 bp    Wild Type
Y695 / Y695 150 bp    Mutant

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2
2.5mM dNTP Mix 0.2mM
100 µMY691 0.5 µM
100 µMY692 0.5 µM
100 µMY695 0.5 µM
100 µMY696 0.5 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
94ºC 0:30
60ºC 0:30 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.