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  • Library prep from single-stranded DNA?

    Hi everyone,

    For an application I'm interested in, I would like to sequence libraries on Illumina (or Solid, I guess) starting from material containing both double and single-stranded DNA. If I'm not mistaken, standard library construction will miss single-stranded DNA, right?

    Is anyone aware of a way to get single- and double- (or at least single- alone) stranded material represented in a library?

    Thanks very much for your help!

  • #2
    Please refer this paper for more on strand specific sequencing

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    • #3
      High-throughput sequencing of cDNA has been used to study eukaryotic transcription on a genome-wide scale to single base pair resolution. In order to compensate for the high ribonuclease activity in bacterial cells, we have devised an equivalent technique optimized for studying complete prokaryotic …

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      • #4
        Thanks to both of you. To clarify, I'm not interested in strand-specific information, as such, but just in generating libraries that would include DNA that starts off as single-stranded.

        Upendra, I'm not sure I see how the paper you posted will help, but maybe I'm just missing it.

        Jos, your reference claims that single-stranded cDNA will make it into a library even if you don't do anything special. The proposed mechanism is through poorly complementary double-stranded regions being processed to ligatable ends during the blunting step. This might well be useful to me, but the paper lacks info about how efficient the process is and what biases it introduces. Does anyone have any insight into this?

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