Stock Number: 006944 Strain Name: B6.129-Mgl1tm1Hed/J

Allele: Mgl1tm1Hed Protocol Name: Mgl1tm1Hed Method: Standard PCR Version: 2.1 Created: 21-November -2008 Updated: 18-November -2009

Expected Results: Wild type = 1450 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8550 - GCG GTT ATT GGA TCT CCT GG - Common oIMR8551 - CTC AGG TTC CCT CTG TGC CAG C - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.66 - 4.66 10 X High Fidelity Buffer 0.80 0.67 X 0.80 50 mM MgSO4 0.60 2.50 mM 0.60 2.5 mM dNTP 0.60 0.13 mM 0.60 20 uM oIMR8550 0.80 1.33 uM 0.80 20 uM oIMR8551 0.80 1.33 uM 0.80 5 M Betaine 1.66 0.69 M 1.66 5 U/ul High Fidelity Platinum Taq 0.08 0.03 U/ul 0.08 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 64 1 min - 4 72 1 min 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 006944 B6.129-Mgl1tm1Hed/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.