Stock Number: 002319 Strain Name: B6.Cg-Tg(BCL2)22Wehi/J

Allele: Tg(BCL2) Protocol Name: Tg(BCL2) Method: Standard PCR Version: 1.1 Created: 02-January -2001 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 170 bp Internal positive control = 324 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 2.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0550 - TGG ATC CAG GAT AAC GGA GG - Transgene oIMR0551 - TGT TGA CTT CAC TTG TGG CC - Transgene 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

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.56 - 2.56 10 X AB PCR BufferII 1.50 1.36 X 1.50 25 mM MgCl2 0.96 2.18 mM 0.96 2.5 mM dNTP 0.96 0.22 mM 0.96 20 uM oIMR0550 0.60 1.09 uM 0.60 20 uM oIMR0551 0.60 1.09 uM 0.60 20 uM oIMR7338 0.48 0.87 uM 0.48 20 uM oIMR7339 0.48 0.87 uM 0.48 5 mM DNA Loading Dye 1.66 0.76 mM 1.66 5 U/ul Taq DNA Polymerase + TaqStart AB (1:1) 0.20 0.02 U/ul 0.20 DNA 1.00 - 1.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 54 30 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 002318 C.Cg-Tg(BCL2)22Wehi/J 002319 B6.Cg-Tg(BCL2)22Wehi/J 002320 B6.Cg-Tg(BCL2)25Wehi/J 002321 B6.Cg-Tg(BCL2)36Wehi/J 002427 C3H/He-Tg(LCKprBCL2)36Sjk/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.