Stock Number: 006879 Strain Name: B6.129-Scd2tm1Myz/J

Allele: Scd2tm1Myz Protocol Name: Scd2tm1Myz Method: Standard PCR Version: 1.1 Created: 02-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 570 bp Heterozygote = 570 bp and 750 bp Wild type = 750

Gel Image

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6981 - GGG CCT GGC TTA CGA CCG GAA GAG AG - - oIMR6982 - ATA GCA GGC ATG CTG GGG AT - - oIMR6983 - GTC ACG AAG TTC CTC AGT TGC CAC - -

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 oIMR6981 0.60 1.00 uM 0.60 20 uM oIMR6982 0.60 1.00 uM 0.60 20 uM oIMR6983 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 69 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 006879 B6.129-Scd2tm1Myz/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.