Stock Number: 007999 Strain Name: B6.129P2-Sorl1Gt(Ex255)Byg/J

Allele: Sorl1Gt(Ex255)Byg Protocol Name: Sorl1Gt(pGT1tfr)1Ucd Method: Standard PCR Version: 1.1 Created: 08-February -2008 Updated: 24-September-2009

Expected Results: Mutant = 681 bp Heterozygote = 364 bp and 681 bp Wild type = 364 bp

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 oIMR7701 - CTC ACT CTT AAG ACC ATG GC - Wild type Forward oIMR7702 - CTT GGC ACC AAA GTC ACT CTA C - Wild type Reverse oIMR7703 - TTA TCG ATG AGC GTG GTG GTT ATG C - Mutant Forward oIMR7704 - GCG CGT ACA TCG GGC AAA TAA TAT C - Mutant Reverse

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 oIMR7701 0.60 1.00 uM 0.60 20 uM oIMR7702 0.60 1.00 uM 0.60 20 uM oIMR7703 0.30 0.50 uM 0.30 20 uM oIMR7704 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 65 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 1 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 007999 B6.129P2-Sorl1Gt(Ex255)Byg/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.