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  • Are the Illumina diagrams correct?

    Is it true that single-stranded DNA libraries are initially BOUND directly to the flow cell as suggested in Illumina's diagrams (see panel b in attached figure). I always assumed they simply hybridized to the primer lawns.


  • #2
    Originally posted by drdna View Post
    Is it true that single-stranded DNA libraries are initially BOUND directly to the flow cell as suggested in Illumina's diagrams (see panel b in attached figure). I always assumed they simply hybridized to the primer
    Yeah, that seems a little misleading. I would say that panel b more accurately represents the state after the initial cycle of amplification (primer extension). The flow cell bound primer has been extended using the hybridized library molecule template, then the original template strand has been denatured and washed away.

    It seems the description of the initial stages in the this thread aren't quite accurate.

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    • #3
      This Illumina figure is also incorrect. As per my understanding, it is not possible to generate double stranded molecules with free ends!



      from the link:

      http://www.illumina.com/content/dam/...troduction.pdf
      Last edited by drdna; 07-20-2015, 07:09 AM.

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      • #4
        Is this illustration better: http://www.illumina.com/documents/pr...sequencing.pdf

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        • #5
          Originally posted by GenoMax View Post
          No, that's the original image that is also inaccurate for a different reason - see original post. It seems that the Illumina folks did not have anyone who understands the technology check the diagrams coming out of their illustration department/provider. I wanted to use an original Illumina illustration in a talk that describes the principle of bridge amplification but the official diagrams will confuse the audience, so I think I'll create my own.

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          • #6
            Originally posted by drdna View Post
            No, that's the original image that is also inaccurate for a different reason - see original post. It seems that the Illumina folks did not have anyone who understands the technology check the diagrams coming out of their illustration department/provider. I wanted to use an original Illumina illustration in a talk that describes the principle of bridge amplification but the official diagrams will confuse the audience, so I think I'll create my own.
            Try the original paper: Nucl. Acids Res. (2000) 28 (20): e87. doi: 10.1093/nar/28.20.e87. Figure 1.

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