Strain Protocols

Strain Information
Strain Code: 01XH4
Strain Name: STOCK Tg(NES-tv-a)J12Ech/Nci
Gene Symbol: tv-a
Protocol Information
Allele: Tg(NES-tv-a)J12Ech
Protocol Number: 1
Introductory Comment: This primer set is used for both the Tg(NES-tv-a)J12Ech and Tg(GFPA-tv-a)10Ech mouse strains. The reaction requires the addition of the additive DMSO along with the use of a heat-activated Taq Polymerase (Platinum Taq from Invitrogen)

Primers

T028 : 5'-cTG cTG ccc GGT AAc GTg Acc-3'
T029 : 5'-Gcc cTG GGG AAG GTc cTG ccc-3'

Product Sizes

Primer Combination Product Size
T028 / T029 477 bp    Transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 1.5 mM
2.5mM dNTP Mix 0.2 mM
DMSO 5%
100 µMT028 0.5 µM
100 µMT029 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
0ºC
94ºC 1:00
65ºC 2:00 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.