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Thread: query regarding the Msh6 exon 1 gene amplification

  1. #1 query regarding the Msh6 exon 1 gene amplification 
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    Dear Sir


    This is Ramesh from cdfd, Hyderabad, India iam working on the HNPCC , iam doing mutational analysis of genes like msh6, msh2,pms2,and mlh1 . i have successfully sequenced many of the samples with those genes and got some satisfied results . But i strucked at the MSH-6 exon1 , because it is pertained with high GC content, i have tried with different primers and sequenced , but no one is matching and the pcr product has very much non specificity appearing on the agarose gel. However we taking the approximate size and purifying them and given for sequencing , but no matched sequence with the reference sequence. for that exon we had ordered almost eight sets of primers to get the amplification, no primer worked on this exon. so please suggest and interpretation on this exon 1 of msh-6.

    so i may go further analysis.

    thanking you

    Ramesh.kamtam
    lmo,cdfd,hyderabad
    India


    Last edited by kamtam ramesh; March 6th, 2013 at 06:44 AM. Reason: to better understanding
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  3. #2  
    New Member
    Join Date
    Mar 2013
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    Dear Sir


    This is Ramesh from cdfd, Hyderabad, India iam working on the HNPCC , iam doing mutational analysis of genes like msh6, msh2,pms2,and mlh1 . i have successfully sequenced many of the samples with those genes and got some satisfied results . But i strucked at the MSH-6 exon1 , because it is pertained with high GC content, i have tried with different primers and sequenced , but no one is matching and the pcr product has very much non specificity appearing on the agarose gel. However we taking the approximate size and purifying them and given for sequencing , but no matched sequence with the reference sequence. for that exon we had ordered almost eight sets of primers to get the amplification, no primer worked on this exon. so please suggest and interpretation on this exon 1 of msh-6.

    so i may go further analysis.

    thanking you

    Ramesh.kamtam
    lmo,cdfd,hyderabad
    India


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