Strain Protocols

Strain Information
Strain Code: 01XH1
Strain Name: B6;129-Dmtf1tm1Cjs/Nci
Gene Symbol: Dmtf1
Protocol Information
Allele: Dmtf1<tm1Cjs>
Protocol Number: 1
Introductory Comment:

Primers

D001 : 5'-GAT GGT ccA ccA TTT AcT TcG-3'
D003 : 5'-GcT GTA GcT GAA AGA GTG GGT Ac-3'
D002 : 5'-AGT ATc cAT cAT GGc TGA TGc-3'

Product Sizes

Primer Combination Product Size
D001 / D003 1400 bp    Wildtype
D002 / D003 1600 bp    Knockout

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.5 mM
2.5mM dNTP Mix 0.2 mM
100 µMD001 0.33 µM
100 µMD003 0.33 µM
100 µMD002 0.33 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
0ºC
94ºC 1:00
65ºC 2:00 35 Cycles
72ºC 1:00
0ºC
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.