Stock Number: 010702 Strain Name: STOCK Gad2tm1(cre/ERT2)Zjh/J

Allele: Gad2tm1(cre/ERT2)Zjh Protocol Name: Gad2tm1(cre/ERT2)Zjh STD PCR Method: Standard PCR Version: 1.0 Created: 23-September-2009 Updated: 20-February -2011

Notes: This assay does not work well without the use of a Hotstart Taq polymerase. Expected Results: Mutant = 247 bp Heterozygote = 247 bp and 507 bp Wild type = 507 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 11400 - ACG TTT CCT GTC CCT GTG TG - Common oIMR3346 - CAG ACG CTG CAG TCT TTC AG - Wild type Reverse oIMR9074 - AGG CAA ATT TTG GTG TAC GG - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.10 - 2.10 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgCl2 0.96 2.00 mM 0.96 DNA 2.00 - 2.00 5 X KAPA LR HotStart Buffer 2.40 1.00 X 2.40 20 uM 11400 0.60 1.00 uM 0.60 20 uM oIMR3346 0.60 1.00 uM 0.60 20 uM oIMR9074 0.60 1.00 uM 0.60 2.5 U/ul KAPA LR HotStart Taq 0.12 0.03 U/ul 0.12

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 60 30sec - 4 72 30sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 010702 STOCK Gad2tm1(cre/ERT2)Zjh/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.