[Question on the sequencing] Details on the carrier sequencing method
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lucventurini commented
Dear authors,
I read today with interest your BMC article on the method implemented in this repository. I am, however, unsure on how it would be replicated experimentally. Specifically, am I correct in understanding that you propose that, in case of a low-concentration sample, the experimenter should "load up" the sample with a known sequence, e.g. the lambda phage used in the article?
If that is the case, I am curious regarding the specific role of the carrier in the reaction. Why would the results be better than without adding the carrier at all? I am afraid that I might have missed the section of the manuscript where you make the argument.
Thank you for your help.
Kind regards
Luca Venturini
CarrCE commented
Hi Luca—
Your description is accurate. It is easy to lose a small amount of DNA
during library preparation, so using a carrier minimizes any such losses
(in addition to maintaining reaction stoichiometry). In addition, nanopore
sequencing with little DNA leads to rapid pore burnout. So the carrier acts
as a pore-maintainer.
Chris
On Thu, Dec 27, 2018 at 8:35 AM Luca Venturini ***@***.***> wrote:
Dear authors,
I read today with interest your BMC article on the method implemented in
this repository. I am, however, unsure on how it would be replicated
experimentally. Specifically, am I correct in understanding that you
propose that, in case of a low-concentration sample, the experimenter
should "load up" the sample with a known sequence, e.g. the lambda phage
used in the article?
If that is the case, I am curious regarding the specific role of the
carrier in the reaction. Why would the results be better than without
adding the carrier at all? I am afraid that I might have missed the section
of the manuscript where you make the argument.
Thank you for your help.
Kind regards
Luca Venturini
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