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Dna chain termination sequencing

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  1. Sep 26, 2012 #1
    1. The problem statement, all variables and given/known data


    A DNA 5-[32P] CCT TCG T TCG TTG TTC CCT A GGC TGT ATA GCC CCT ACC TTT TTG GTA GGG GCT ATA CAG CC was elongated by DNA polymerase in four reaction mixtures in the presence of dATP, dTTP, dGTP (dCTP was omitted due to an experimental mistake) and one ddNTPs. The four reaction mixtures contained different ddNTPs. Predict the band pattern after separation of the reaction mixtures ion gel and autoradiography.


    I thought I was starting to understand until I saw a classmate talking about the primer ending at the lone A in the middle.

    I thought that you would start sequencing at the 3 prime end, so you would get GGC ..etc.
    but since there are no C;s since dCTP was omitted, it would just stop there, so you would just get a band patter of two lines in the ddGTP column.

    But now I don't get this whole primer thing. Is that entire chunk from that single A in the middle all the way to the end just a primer?

    So then would I sequence,
    AGGGAAC and then stop at that C?

    Is this right? And if so, how do you know that the primer ends at that middle A?


    2. Relevant equations



    3. The attempt at a solution
     
  2. jcsd
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