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    Hi
    Does anyone have a simple step my step method for Sanger Sequencing for AQA BIOL 5 ?
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    From my notes:

    1) To four separate tubes add:
    -single stranded DNA template (sample)
    -DNA polymerase
    -DNA (radioactively/fluorescently) labelled primers
    -Excess free nucleotides
    -1 type of modified nucleotide

    2) Allow tubes to undergo PCR - many copies of DNA strands formed, of various lengths (as each modified nucleotide terminates strand synthesis at different points randomly)

    3) Separate fragments (one tube per well) using gel electrophoresis

    4) Visualise separation using UV/X-Rays (primers were labelled so we can do this)

    5) Smallest fragment moves down to the bottom so read up nucleotide by nucleotide - forming the sequence of bases complementary to original strand
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    (Original post by Serine Soul)
    From my notes:

    1) To four separate tubes add:
    -single stranded DNA template (sample)
    -DNA polymerase
    -DNA (radioactively/fluorescently) labelled primers
    -Excess free nucleotides
    -1 type of modified nucleotide

    2) Allow tubes to undergo PCR - many copies of DNA strands formed, of various lengths (as each modified nucleotide terminates strand synthesis at different points randomly)

    3) Separate fragments (one tube per well) using gel electrophoresis

    4) Visualise separation using UV/X-Rays (primers were labelled so we can do this)

    5) Smallest fragment moves down to the bottom so read up nucleotide by nucleotide - forming the sequence of bases complementary to original strand
    Thank You !
 
 
 
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