Intro to Sequencing by Synthesis: Industry-leading Data Quality

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This video provides an overview of the DNA sequencing workflow on an Illumina sequencer. The process includes cluster generation on a system flow cell, sequencing with Illumina’s proprietary sequencing by synthesis technology, and data analysis.

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Comment (57)

  1. Hello, nice video, I've always wondered how can the image software to know the identity of each new base added to the growing strand, given that this chain is composed by a mixture of many fluorescent nucleotids?

  2. Nice video, though I would like to know the method by which the cleavage process before the addition of labeled nucleotides affects only the reverse strands and not the forward ones, given that both of them have identical adapters.

  3. Can someone explain to me why we need 2 reads? Also, do you use barcoding when analyzing self "made" (PCR) amplicons generated from bacterial population 16s rRNA Gene fragments? Or just when you get a sample from a library and such?

  4. From the graphic view of mapped reads, if reverse reads (generally shown in RED) were shown mapping to the reference sequence then can we say that reads are coming from antisense strand ?

  5. Could someone explain me one thing? Each cluster contains milions of copies of the same fragments or for example cluster 1 have milions of copies of fragment 1 and cluster 2 milions of copies of fragment 2? I think the second one but i'm not so sure

  6. it doesnt appear to be explained why the reverse sequence is also sought. i was expecting at the end for them to say "and the reverse strand sequence is used to verify the original forward sequence". if not for this reason, then why?

  7. I have a doubt on this, all the flow cell adapters have the same sequence? Or every well have their own adapters for that cluster? In the first case, how is controlled that only one sequence binds to a given cluster, and not a mix of differents sequences bind in the same cluster?

  8. Hi, I have a few questions that need urgent answers:
    1/ How flourescent label in the blocking group are removed from the nucleotide?
    2/ How to wash away free nucleotide?
    3/ Why do we have to sequece 2 strain of DNA?
    4/ What is the function of index 1 and 2 in adapter attached to DNA?
    Thank you.

  9. read1 product washed away
    index1 primer attatched

    question) why is this running individually?
    why is index 1 primer product generate newly?

  10. read 1 and read 2 give the same result but thet are done in opposite directions. Just so you know and don't have to waste hours trying to puzzle together little details that don't make sense at first.

  11. I have a question about this process. There are two types of oligos fixed in the surface and two types of adaptors in the strand DNA, so one adapter is complementary to one oligo and the other adapter is equal to the second oligo (and consequently different to the first oligo)?


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