DNA Sequencing

DNA Sequencer

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11 Comments

1.   IloveYOUviruses   |  Wednesday, 21 July 2010 at 7:22 pm

   WHAT!? DNA in a conveyor belt????? AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA(must be telekinesis) -_-

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2.   Dava1717   |  Wednesday, 21 July 2010 at 8:05 pm

   Can anyone explain the Maxam-Gilbert Method?

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3.   shir312   |  Wednesday, 21 July 2010 at 8:20 pm

   Because different ddNTPs attach to the dna strand in different places, one will randomly attach right after the primer on one piece, another will randomly attach two nucleotides after the primer, and so on. Then using gel electrophoresis, you can separate the strands by size and record the ending ddNTPs, as shown above, to give you the full sequence of the DNA strand.

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4.   marcianos   |  Wednesday, 21 July 2010 at 9:09 pm

   Up to what I know (and as you can see in the video), you can run all of them in the same tube as long as each ddNtP has a different fluorescent mark. If They all have the same mark, then you need to run them in different tubes.

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5.   bruckwine1978   |  Wednesday, 21 July 2010 at 9:12 pm

   ya mark just like that 4 separate tubes for each base to read one sequence

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6.   bruckwine1978   |  Wednesday, 21 July 2010 at 9:49 pm

   Yea that’s right mark – so they need to run all 4 rxns in 4 separate tubes to read one sequence just like that.

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7.   markgg1   |  Wednesday, 21 July 2010 at 10:27 pm

   Thanks!

   its just clicked in my mind how it works I think

   E.g. for length 1 bases, the end might be A

   2 bases would end T

   3 bases end C

   4 bases end G

   So, it would read ATCG?

   Am I right?

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8.   adnansaami   |  Wednesday, 21 July 2010 at 10:55 pm

   does anyone have a flow diagran of how to sequence dna using whole blood using the sanger method

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9.   bruckwine1978   |  Wednesday, 21 July 2010 at 11:33 pm

   In reality they use all FOUR ddNTPs at the same time, e.g. ddATP, ddTTP, ddCTP, ddGTP, on each sample – so you get fragemnts that end at each letter throughout the sample..based on lengths you can tell which base is at which position.

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10.   meita01   |  Thursday, 22 July 2010 at 12:30 am

    how can an accurate catalogue of a DNA sequence be created by only looking at the end ddNTP base?

    For instance, if you have a 5-base stand where the 5th base is the ddNTP and this is sequenced how can you find out about the other 4 bases that come before the ddNTP???

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11.   befz88   |  Thursday, 22 July 2010 at 12:43 am

    ya okay, but it doesnt explain the whole sequencing process.

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