Stock Number:002020
Strain Name:C57BL/6J-ApcMin/J
ApcMin
ApcMin
End Point Analysis
4.1
15-September-2008 20-November -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
Sequence: GCCATCCCTTCACGTTAGGAAACAGAAAGCTCTAGAAGCTGAGCTAGATGCTCA
GCATTTATCAGAAACCTTCGACAACATTGACAACCTAAGTCCCAAGGCCTCTCAC
CGGAGTAAGCAGAGACACAAGCAGAATCTTTATGGTGACTATGCTTTTGACGCCA
ATCGACATGATGATAGTAGGTCAGACAATTTCAATACTGGAAACATGACTGTTCTT
TCACCATATTTAAATACTACGGTATTGCCCAGCTCTTCTTCCTCAAGGGGAAGTTTA
GACAGTTCTCGTTCTGAGAAAGACAGAAGTT(t/a)GGAGAGAGAGCGAGGTATTGG
CCTCAGTGCTTACCATCCAACAACAGAAAATGCAGGAACCTCATCAAAACGAGGT
CTGCAGATCACTACCACTGCAGCCCAGATAGCCAAAGTTATGGAAGAAGTATCAG
CCATTCATACCTCCCAGGACGACAGAAGTTCTGCTTCTACCACCGAGTTCCATTGT
GTGGCAGACGACAGGAGTGCGGCACGAAGAAGCTCTGCCTCCCACACACACTCA
AACACATACAACTTCACTAAGTCGGAAAATTCAAATAGGACATGCTCTATGCCTTAT
GCCAAAGTGGAA
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
oIMR3200 - GAA GGT GAC CAA GTT CAT GCT TCC AAT ACC TCG CTC TCT CTC CA - Wild type
oIMR3201 - GAA GGT CGG AGT CAA CGG ATT CCA ATA CCT CGC TCT CTC TCC TA - Mutant
oIMR3202 - GCC CAG CTC TTC TTC CTC AA - Common
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 6.60 - 6.60
20 X Ampliflour SNP FAM 0.50 0.91 X 0.50
20 X SNP JOE 0.50 0.91 X 0.50
20 X Primer Mix 0.50 0.91 X 0.50
10 X Rxn Mix S+ 1.00 0.91 X 1.00
2.5 mM dNTP 0.80 0.18 mM 0.80
5 U/ul Titanium Taq Pol. 0.10 0.05 U/ul 0.10
DNA 1.00 - 1.00
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
002020 C57BL/6J-ApcMin/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.