Strain Protocols

Strain Information
Strain Code: 01X62
Strain Name: B6.129-Cdkn2atm4Cjs/Nci
Gene Symbol: Cdkn2a
Protocol Information
Allele: Cdkn2a<tm4Cjs>
Protocol Number: 1
Introductory Comment: This protocol use only the SpF/R primers to confirm presence/absence of LOX sites at the Sp (Spel) sites

Primers

SPF : 5'-TACTGCAGCCAGACCACTAGG-3'
SPR : 5'-CGGAGATTGAGAAAGCGG-3'

Product Sizes

Primer Combination Product Size
SPF / SPR WT    263BP
SPF / SPR LoxP    360 BP

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2 mM
2.5mM dNTP Mix 0.2mM
100 µMSPF 0.5 µM
100 µMSPR 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.