Strain Protocols

Strain Information
Strain Code: 01XJA
Strain Name: B6.129S7-Pmltm1Ppp/Nci
Gene Symbol: PML
Protocol Information
Allele: Pml<tm1Ppp>
Protocol Number: 1
Introductory Comment:

Primers

Y317 : 5'-TTT CAG TTT CTG CGC TGC C-3'
Y319 : 5'-TTG GAC TTG CGC GTA CTG TC-3'
Y318 : 5'-CGA CCA CCA AGC GAA ACA-3'

Product Sizes

Primer Combination Product Size
Y317 / Y319 450bp    Wild type
Y318 / Y319 650bp    Knockout

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMY317 0.5 µM
100 µMY319 0.5 µM
100 µMY318 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.