Stock Number: 006900 Strain Name: B6.129-St3gal4tm1.1Jxm/J

Allele: St3gal4tm1.1Jxm Protocol Name: St3gal4tm1.1Jxm MCA Method: Melt Curve Analysis Version: 1.0 Created: 20-November -2009 Updated: 20-November -2009

Notes: Melting curve analysis is done using a Roche Light Cycler 480. Expected Results: Het = 83C +/- 1.0 area 6 Het2 = 87C +/- 1.0 area 5 WT = 83C +/- 1.0 area 10

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6890 - GAC GCC ATC CAC CTA TGA G - - oIMR6891 - GGC TGC TCC CAT TCC ACT - - oIMR6892 - GGC TCT TTG TGG GAC CAT CAG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.20 - 2.20 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR6890 0.60 1.00 uM 0.60 20 uM oIMR6891 0.60 1.00 uM 0.60 20 uM oIMR6892 0.60 1.00 uM 0.60 DNA 2.00 0.00 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 65 1 min - 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 006900 B6.129-St3gal4tm1.1Jxm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.