Stock Number: 008217 Strain Name: B6.129S1-Car4tm1Sly/J

Allele: Car4tm1Sly Protocol Name: Car4tm1Sly Method: Standard PCR Version: 2.0 Created: 11-August -2009 Updated: 24-September-2009

Expected Results: HET = 700 bp and 548 bp MUT = 700 bp WT = 548 bp

Gel Image

no data found

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8222 - TGT GGA TCT TGA TGG GTT GTT TGT ACA CAG TCC - - oIMR8223 - TGA GGT CAA TTC CCA GCT CCC ACA TTG TTG C - - oIMR8970 - GGC CTG AGA AGT GGC CTG GA - - oIMR8971 - AGC CCT GTC CTG GGC CTG AT - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.26 - 3.26 10 X PCR Buffer II (ABI) 0.80 0.67 X 0.80 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8222 0.80 1.33 uM 0.80 DNA 2.00 - 2.00 20 uM oIMR8970 0.40 0.67 uM 0.40 20 uM oIMR8971 0.40 0.67 uM 0.40 20 uM oIMR8223 0.80 1.33 uM 0.80 5 M Betaine 1.50 0.63 M 1.50 5 U/ul Taq DNA Polymerase + TaqStart AB (1:1) 0.12 0.05 U/ul 0.12

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 68 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold**

Reaction/Components  None found. This genotyping protocol is used for the following strains:   Stock Number Strain Name 008217 B6.129S1-Car4tm1Sly/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  None found.