Stock Number: 000219 Strain Name: C57BL/6J-Slc30a4lm/J

Allele: Slc30a4lm Protocol Name: Slc30a4lm Method: End Point Analysis Version: 2.1 Created: 26-September-2008 Updated: 24-September-2009

Notes: End point analysis is done using Amplifluor SNPS Genotyping System from Millipore. End point analysis protocols are run on an ABI 7000, 7500, or the Roche LightCycler 480 Expected Results: Mutant = red Wild type = green

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR3450 - TCC TTC ACA GTG TCA CTC AGA T - Common oIMR4229 - GAA GGT CGG AGT CAA CGG ATT GTG TGC TTA TAG CTG CAT ACA TCA TAC - - oIMR4230 - GAA GGT GAC CAA GTT CAT GCT GTG TGC TTA TAG CTG CAT ACA TCA TAT - - oIMR4231 - GCC TGG CTG TCA ACT AGC TTT T - Common

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.60 - 4.60 20 X Ampliflour SNP FAM 0.50 1.00 X 0.50 20 X SNP JOE 0.50 1.00 X 0.50 20 X Primer Mix 0.50 1.00 X 0.50 10 X Rxn Mix S+ 1.00 1.00 X 1.00 2.5 mM dNTP 0.80 0.20 mM 0.80 5 U/ul Titanium Taq Pol. 0.10 0.05 U/ul 0.10 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 95 5 min - 2 94 30 sec - 3 55 30 sec - 4 72 40 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 000219 C57BL/6J-Slc30a4lm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.