Strain Protocols

Strain Information
Strain Code: 01XT3
Strain Name: FVB.Cg-Tg(KRT14-HPV16)wt1Dh/Nci
Gene Symbol: HPV16
Protocol Information
Allele: Tg(KRT14-HPV16)wt1Dh
Protocol Number: 1
Introductory Comment: This line contains the HPV16 early coding region controlled by the KRT14 promoter. Primers directed at the HPV16 element distinguish this line from a wt line, which does not contain an HPV16 element.

Primers

JA7 : 5'-aga act gca atg ttt cag gac cca cag-3'
JA4 : 5'-tct gca aca aga cat aca tcg acc gg-3'
Diagram showing the primer relataionships for Strain 01XT3.  This line contains the HPV16 early coding region controlled by the KRT14 promoter.  Primers directed at the HPV16 element distinguish this line from a wt line, which does not contain an HPV16 element.

Product Sizes

Primer Combination Product Size
JA7 / JA4 431bp    Tg

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0mM
2.5mM dNTP Mix 0.2mM
100 µMJA7 0.5 µM
100 µMJA4 0.5 µM
5U/┬Ál Taq Pol 0.015 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 0:30
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.