4 thoughts on “G20 – Sprason

  1. Is the nanopore techonolgy very reliable? Does it make mistakes? Could you also explain what the dots in the blast result represent? I’m still a little confused on what is is showing, and what the dots tell you.

    1. Hi Madeline, from this work we’ve currently found that nanopore is almost as accurate as illumina, up to 99.99%. However, since most of our work was looking at previously un-sequenced phages, we can’t say at this moment how reliable that accuracy number is, because we only have 1 illumina sequenced phage to base our results off of.
      Regarding the BLAST results, they were used to generate reference sequences/positive controls which I could compare the nano-pore sequenced/new phages to (as theyve not been sequenced by illumina yet). I then used the new sequences and the BLAST results to make a dotplot. Dotplots show how similar sequences are to each other. In the first, you can see that the lines are discontinuous which suggested something wasn’t quite right. After i adjusted the start sites you can see the 2nd dot plot looks a lot better with very straight diagonal lines. This means that the new phages were very close to their BLAST references, in other words, they were accurately sequenced by nanopore.

  2. Since you mentioned nanopore technology is inexpensive and accessible enough for the classroom does that mean it’s approximate but not highly accurate in sequencing?

    1. Hi Vanshita, Not at all! Nanopore is far cheaper and accessible than illumina as it only requires a small USB sized device connected to a computer rather than a huge, expensive machine. It is also highly accurate, from this work we predict it is 99.99% accurate compared to illumina’s 99.999%. Admittedly, this degree less of accuracy does mean that when sequencing very large sequences (say an organisms with multiple chromosomes and billions of base pairs, compared to a phage which is a virus with closer to 60,000 base pairs) that nanopore is not as effective, but it can still produce very valid sequencing data.

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