Stock Number: 006412 Strain Name: B6.129-Il12btm1Lky/J

Allele: Il12btm1Lky Protocol Name: Il12btm1Lky STD PCR Method: Separated PCR Version: 3.0 Created: 02-March -2012 Updated: 02-March -2012

Notes: This assay does not work well without the use of a Hotstart. Expected Results: Mutant = 300 bp Heterozygote = 300 bp and 468 bp Wild type = 468 bp

Gel Image

no data found

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6522 - GAC CCA GTG TCT GGG GAT AA - - oIMR6523 - TTG CAT AAT AGG GCC TGG TC - - oIMR8292 - CCA AAA GAC GGC AAT ATG GT - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.55 - 4.55 5 X Kapa 2G HS buffer 2.40 1.00 X 2.40 25 mM MgCl2 0.96 2.00 mM 0.96 10 mM dNTPS-kapa 0.24 0.20 mM 0.24 20 uM oIMR6522 0.30 0.25 uM 0.30 20 uM oIMR6523 0.30 0.25 uM 0.30 5 mM 10x Loading Dye 1.20 0.50 mM 1.20 2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 se - 3 59 1 min - 4 72 30 se - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.55 - 4.55 5 X Kapa 2G HS buffer 2.40 1.00 X 2.40 25 mM MgCl2 0.96 2.00 mM 0.96 10 mM dNTPS-kapa 0.24 0.20 mM 0.24 20 uM oIMR6522 0.30 0.25 uM 0.30 20 uM oIMR8292 0.30 0.25 uM 0.30 5 mM 10x Loading Dye 1.20 0.50 mM 1.20 2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05 DNA 2.00 - 2.00 This genotyping protocol is used for the following strains:   Stock Number Strain Name 006412 B6.129-Il12btm1Lky/J 009618 NOD.129(B6)-Il12btm1Lky/JbsJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  None found.