Strain Protocols

Strain Information
Strain Code: 01XMD
Strain Name: B6;129S-Mdm4GT(VICTR20)7Lex Mdm2tm1BayTrp53tm1Brd/Nci
Gene Symbol: Mdm4
Protocol Information
Allele: Gt(VICTR20)7Lex
Protocol Number: 1
Introductory Comment: MdmX (4) Genotyping Protocol

Primers

Y846 : 5'-tag gtt caa ttc ctg cag ccc-3'
Y847 : 5'-gag aat ttc agt tca gtg tga gt -3'
Y848 : 5'-gcc ttg caa aat ggc gtt act t-3'

Product Sizes

Primer Combination Product Size
Y846 / Y847 350 bp    Wild Type
Y848 / Y847 250 bp    Mutant

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2
2.5mM dNTP Mix 0.2mM
100 µMY846 0.5 µM
100 µMY847 0.5 µM
100 µMY848 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 99:99

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.