Strain Protocols

Strain Information
Strain Code: 01XT4
Strain Name: FVB.Cg-Tg(Dct-rtTA2S*M2)#Mrln Tg(tetO-HIST1H2BJ/GFP)47Efu/Nci
Gene Symbol: rtTA
Protocol Information
Allele: Tg(Dct-rtTA2S*M2)#Mrln
Protocol Number: 1
Introductory Comment: The iDct-GFP mice (01XT4) contain two transgenes, so two genotyping PCR reactions need to be run for each mouse. The genotyping PCR protocol 1 is used to type the Dct-rtTA gene and protocol 2 is forTRE-H2BGFP gene.

Primers

Y104 : 5'-ACT AAG TAA GGA TCA ATT CAG-3'
Y105 : 5'-TGT ACT AGG CAG ACT GTG-3'

Product Sizes

Primer Combination Product Size
Y104 / Y105 370 bp    transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2 mM
2.5mM dNTP Mix 0.2mM
100 µMY104 0.5 µM
100 µMY105 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
55ºC 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.