Stock Number: 008120 Strain Name: B6.129S4-Timp2tm1Pds/J

Allele: Timp2tm1Pds Protocol Name: Timp2tm1Pds Method: Standard PCR Version: 3.0 Created: 30-July -2009 Updated: 24-September-2009

Notes: TGS design to 3' Smal insertion site w/Pgkneo cassette (reverse orientation).  KJ 12/08 Expected Results: HET = 250 bp and 283 bp MUT = 250 bp WT = 283 bp

Gel Image

no data found

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR7415 - GCC AGA GGC CAC TTG TGT AG - Mutant oIMR9312 - TTC TTT TCC CCT CTG AGC TG - Wild type Forward oIMR9313 - CTG CCC TGC TAA GCA AAA AC - Common

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 20 uM oIMR7415 0.60 1.00 uM 0.60 5 U/ul Platinum Taq Pol. 0.08 0.03 U/ul 0.08 20 uM oIMR9312 0.60 1.00 uM 0.60 20 uM oIMR9313 0.60 1.00 uM 0.60 DNA 2.00 - 2.00 ddH2O 3.34 - 3.34 2.5 mM dNTP 0.96 0.20 mM 0.96 10 X Platinum Buffer 1.20 1.00 X 1.20 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgSO4 0.96 2.00 mM 0.96

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 62 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 008120 B6.129S4-Timp2tm1Pds/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.