Strain Protocols

Strain Information
Strain Code: 01CMM
Strain Name: C57BL/6-Mir10btm1.2Arte/Nci
Gene Symbol: miR-10b
Protocol Information
Allele: Mir10b<tm1.2Arte>
Protocol Number: 1
Introductory Comment:

Primers

Y781 : 5'-CAG GCT TGG TGA GTT CCT G-3'
Y782 : 5'-TGT CTT CTT CCC ACA CAA GG-3'

Product Sizes

Primer Combination Product Size
Y781 / Y782 514 bp    Wildtype
Y781 / Y782 203 bp    Knockout

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2 mM
2.5mM dNTP Mix 0.2mM
100 µMY781 0.5 µM
100 µMY782 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.