Stock Number: 014579 Strain Name: B6.NOD-Tg(Foxp3-EGFP/cre)1aJbs/J

Allele: Dicer1tm1Bdh Protocol Name: Dicer1tm1Bdh Method: Standard PCR Version: 2.0 Created: 23-April -2010 Updated: 22-August -2012

Expected Results: Mutant = 420 bp Heterozygote = 351 bp and 420 bp Wild type = 351 bp

Gel Image

Image Image file View Full Size

Gel Information: Separated by gel electrophoresis on a 3% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6305 - CCT GAC AGT GAC GGT CCA AAG - - oIMR6569 - CAT GAC TCT TCA ACT CAA ACT - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.92 - 4.92 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR6305 0.60 1.00 uM 0.60 20 uM oIMR6569 0.60 1.00 uM 0.60 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul KAPATaq 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 55 30 sec - 4 72 30 sec repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 006001 STOCK Dicer1tm1Bdh/J 014579 B6.NOD-Tg(Foxp3-EGFP/cre)1aJbs/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.