Stock Number: 012583 Strain Name: B6.129P2(Cg)-Hprttm68(Ple127-lacZ)Ems/Mmjax

Allele: Hprttm68(Ple127-lacZ)Ems Protocol Name: Hprttm68(Ple127-lacZ)Ems Method: Standard PCR Version: 1.0 Created: 04-March -2010 Updated: 27-May -2010

Notes: Mutant is genotyped by the generic lacZ assay CRB 5-27-10 Expected Results: Wt = 371 bp

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR9435 - AGA TGA GCT TGG TTG CTG GAG TG - Wild type Forward oIMR9436 - TGC TGG ACA TCC CTA CTA ACC CA - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2.5 mM dNTP 0.96 0.20 mM 0.96 DNA 2.00 0.00 2.00 ddH2O 4.92 - 4.92 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 20 uM oIMR9435 0.60 1.00 uM 0.60 20 uM oIMR9436 0.60 1.00 uM 0.60 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 64 30 sec - 4 72 30 sec - 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 012583 B6.129P2(Cg)-Hprttm68(Ple127-lacZ)Ems/Mmjax Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.