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If one of the dideoxynucleotides is omitted while using the Sanger method how would that affect the chain termination method of DNA sequencing?

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  • $\begingroup$ Are you talking that you don't put in a whole base (like T) or if you have a deletion? $\endgroup$
    – SolarLunix
    Commented Sep 13, 2015 at 11:44
  • $\begingroup$ if by one of the dideoxynucleotides, you mean one of ddatp, or.ddctp, ddgtp, or ddttp was ommited from the process, then the sequencing would only be able to terminate for the other three dideoxynucleotides. And you would not be able to get the full sequence, and only nucleotides of whatever three you had. $\endgroup$
    – Ro Siv
    Commented Sep 13, 2015 at 11:45
  • $\begingroup$ @RoSiv if you have an answer then post that as an answer. Comments should be used only to seek clarification or suggest improvements. $\endgroup$
    – WYSIWYG
    Commented Sep 13, 2015 at 17:57
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    $\begingroup$ @user18817 you would possibly benefit from familiarizing yourself with the BiologySE homework policy: biology.stackexchange.com/help/homework $\endgroup$
    – mdperry
    Commented Sep 13, 2015 at 23:15
  • $\begingroup$ @WYSIWYG I was not sure if I was right. I am only an undergrad and I dont wanna give out a wrong answer to someone. Sorry. $\endgroup$
    – Ro Siv
    Commented Sep 13, 2015 at 23:30

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If by one of the dideoxynucleotides, you mean one of ddATP, ddCTP, ddGTP, or ddTTP was omitted from the process, then the sequencing would only be able to terminate for the other three dideoxynucleotides. And you would not be able to get the full sequence, and only nucleotides of whatever three you had.

The function of ddNTPs (the dideoxnucleotides) is to stop the elongation of DNA and to subsequently be read by either fluorescence or radioactivity.

Source: https://en.wikipedia.org/wiki/Sanger_sequencing

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  • $\begingroup$ Note that in Sanger sequencing the ddNTPs are not added together in the same mix. If you omit one ddNTP then you won't be able to sequence that particular base in the DNA. Though you may be able to infer the 4th base from the other three, the confidence of the read would be low. $\endgroup$
    – WYSIWYG
    Commented Sep 14, 2015 at 12:08

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