Stock Number:014093
Strain Name:STOCK Tg(tetO-CHRM3*)1Blr/J
** human CHRM3*Y149C
** human CHRM3*Y149C
Pyrosequencing
1.0
30-November -2011 16-December -2011
Notes:

This assay will NOT distinguish hemizygous from homozygous trangenic animals.  This assay only confirms the point mutation.

This genotyping assay uses pyrosequencing technology and is run on the Biotage PSQ 96MA. The Jackson Laboratory is not posting the complete details of our pyrosequencing genotyping assays as the primers for pyrosequencing cannot be used for sequencing using more traditional methods. The wild type and mutant nucleotides and the flanking DNA sequence are provided below.
Expected Results:
Mutant = C/C
Wild type has no PCR product for pyrosequencing.
 
GCCATTGACTa/gCGTAGCCAG
 
Sequencing primer anneals to the reverse strand.
Sequence:

 

TCATCGGCAACATCCTGGTAATTGTGTCATTTAAGGTCAACAAGCAGCTGAAG
ACGGTCAACAACTACTTCCTCTTAAGCCTGGCCTGTGCCGATCTGATTATCGGG
GTCATTTCAATGAATCTGTTTACGACCTACATCATCATGAATCGATGGGCCTTAG
GGAACTTGGCCTGTGACCTCTGGCTTGCCATTGACT(a/g)CGTAGCCAGCAATGCC
TCTGTTATGAATCTTCTGGTCATCAGCTTTGACAGATACTTTTCCATCACGAGG
CCGCTCACGTACCGAGCCAAACGAACAACAAAGAGAGCCGGTGTGATGATCG
GTCTGGCTTGGGTCATCTCCTTTGTCCTTTGGGCTCCTGCCATCTTGTTCTGGC
AATACTTTGTTGGAAAGAGAACTGTGCCTCC

 
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
014093 STOCK Tg(tetO-CHRM3*)1Blr/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.