Stock Number:005657
Strain Name:B6.FVB(129)-Tg(Myh6-cre/Esr1*)1Jmk/J
Tg(Myh6-cre/Esr1*)1Jmk
Tg(cre/Esr1)
Standard PCR
1.1
08-July -2005 24-September-2009
Notes:

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

Customers report that the annealing temperature of 54 C works well.  70 C annealing did not work for them.
Expected Results:

Transgene = ~440 bp
Internal positive control = 324 bp
Gel Image 
Image Image file
View Full Size
Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
oIMR3798 - AGG TGG ACC TGA TCA TGG AG - Transgene Reverse
oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward
oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse
oIMR8346 - ATA CCG GAG ATC ATG CAA GC - Transgene Forward
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 3.12 - 3.12
10 X AB PCR BufferII 1.20 1.00 X 1.20
25 mM MgCl2 1.20 2.50 mM 1.20
2.5 mM dNTP 0.96 0.20 mM 0.96
20 uM oIMR8346 0.60 1.00 uM 0.60
20 uM oIMR3798 0.60 1.00 uM 0.60
20 uM oIMR7338 0.30 0.50 uM 0.30
20 uM oIMR7339 0.30 0.50 uM 0.30
5 mM DNA Loading Dye 1.66 0.69 mM 1.66
5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 3 min -
2 94 30 sec -
3 70 45 sec -
4 72 45 sec repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
005650 STOCK Tg(Myh6-cre/Esr1*)1Jmk/J
005657 B6.FVB(129)-Tg(Myh6-cre/Esr1*)1Jmk/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.