Stock Number: 008656 Strain Name: B6.Cg-KitlSl-gb/MbeJ

Allele: KitlSl-gb Protocol Name: Cg-KitlSl-gb Method: Standard PCR Version: 2.0 Created: 07-August -2009 Updated: 24-September-2009

Expected Results: HET = 175 bp and 208 bp MUT = 208 bp WT = 175 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR9456 - AGT TCA GTC ATA GAT TGG AG - - oIMR9457 - TGT ATC AAA AGG GTC GGG AC - - oIMR9458 - ATT GCT GTA CTT GCT GCC TG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR9456 0.60 1.00 uM 0.60 20 uM oIMR9457 0.60 1.00 uM 0.60 20 uM oIMR9458 0.60 1.00 uM 0.60 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 54 1 min - 4 72 1 min 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 008656 B6.Cg-KitlSl-gb/MbeJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.