Stock Number: 009603 Strain Name: B6.129S4-Kcnn3tm1Jpad/J

Allele: Kcnn3tm1Jpad Protocol Name: Kcnn3tm1Jpad STD PCR Method: Standard PCR Version: 1.0 Created: 16-May -2009 Updated: 09-August -2010

Expected Results: Mutant =140 bp Heterozygote = 140 bp and 432 bp Wild type = 432 bp

Gel Image

no data found

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 8461 - GGA GTA CTC ACC CCA ACA GC - Mutant Reverse 8462 - AGA GGA CTG GAG CCA AGA CA - Common 8463 - ACA GAC CAG GAT GGA CAA GC - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 10 X AB PCR BufferII 1.20 1.00 X 1.20 ddH2O 3.90 - 3.90 5 U/ul Taq DNA Polymerase 0.12 0.05 U/ul 0.12 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 20 uM 8461 0.20 0.33 uM 0.20 20 uM 8462 0.40 0.67 uM 0.40 20 uM 8463 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 68 1 min - 4 72 1 min - 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 009603 B6.129S4-Kcnn3tm1Jpad/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.