Strain Protocols

Strain Information
Strain Code: 01XE5
Strain Name: FVB/N-Tg(Trp53R172H)8512Jmr/Nci
Gene Symbol: Trp53
Protocol Information
Allele: Tg(Trp53R172H)8512Jmr
Protocol Number: 1
Introductory Comment: This strain contains a transgene derived of the R172H mutant variant of the Trp53 gene driven by the WAP promoter. The genotying strategy is to detect this transgene with primer T022, which is specific for the WAP promoter, and T023, which is specific for Exon 2 of Trp53.

Primers

T022 : 5'-ccg tcg acg gcc aca gtg aag acc tcc ggc cag-3'
T023 : 5'-gcc tga aaa tgt ctc ctg gct cag agg g-3'
Diagram showing the primer relataionships for Strain 01XE5.  This strain contains a transgene derived of the R172H mutant variant of the Trp53 gene driven by the WAP promoter.  The genotying strategy is to detect this transgene with primer T022, which is specific for the WAP promoter, and T023, which is specific for Exon 2 of Trp53.

Product Sizes

Primer Combination Product Size
T022 / T023 ~1300bp    Transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 1.5mM
2.5mM dNTP Mix 0.2 mM
DMSO 2%
100 µMT022 0.5 µM
100 µMT023 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 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.