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  • GenoMax
    replied
    Originally posted by TonyBrooks View Post
    Maybe ONT have learnt from PacBio's mistakes. Remember the anticlimax when you first saw PacBio data?
    There is some truth to that. Unfortunately this is starting to feel like "Duke Nukem Forever".

    Leave a comment:


  • TonyBrooks
    replied
    Maybe ONT have learnt from PacBio's mistakes. Remember the anticlimax when you first saw PacBio data?

    Nanopore error rate is around 4% ~Q14, or at least it was 9 months ago.
    The problem is we've all been spoilt by Illumina SBS and will see anything with < Q30 (or even Q20). They've gone on record saying they won't release anything until error is <1% (which is probably nearer the acceptable level). How far away from that they are is anyone's guess at the moment.

    Leave a comment:


  • Nanoporous
    replied
    Originally posted by ECO View Post
    Overall my conversation was pretty disappointing as far as new info (although the gentleman I talked to resisted a pretty good grilling from me and a 454/Roche product manager).

    Points from my conversation that I can remember...
    • GridION == "4-5x more sensors" than the MinION (no comparative runtime specs).
    • GridION can sip from a 96 well plate full of samples (this was demo'd as a flat bottom black microtiter plate with no septa...hmmm).
    • Acquisition speed == "bases per second".
    • The above picture suggesting sample prep is just a "10bp overhang" is not correct and not confirmed
    • The prototypes of the MinION ended up much larger than shown at AGBT due to heat issues (had to "add fans").
    • They will not confirm which gating protein (exo or phi29) or which pore (mspA or a-HL), but that the gate protein is not covalently coupled to the pore. (which strongly suggests to me that it is phi29...the exo-release method would undoubtedly need exquisite positioning for efficient detection)
    • According to the CEO, the DNA input concentration is "nanomolar". This makes sense to be given the binding kinetics of phi29 to it's binding site...but translates into high concentrations for a single molecule platform. When questioned about the possibility of doing very dilute samples where all the molecules matter, he suggested some method of on-chip enrichment/concentration (?????).
    • They are cagey about data release because they want to (my paraphasing) "release data when it's ready"

    Anyway, that's all I got. The ratio of ([booth girls] + [Apple-esque brushed aluminum] + [blinky lights]) to (data) was undefined...so that seems to speak for itself.
    Thanks for the very nice summary.

    For the 'gate' protein, I think they are using some sort of a motor protein to slow down the motion of the DNA in a stepwise manner. As per Clive Brown's talk at AGBT, it is certainly not attached to the pore, and that it is definitely not phi29 (which is what the Akeson and Gundlach groups are using). A polymerase like that would require that you add NTPs. So probably a helicase or something.
    Last edited by Nanoporous; 11-08-2012, 03:27 PM.

    Leave a comment:


  • HESmith
    replied
    Originally posted by ECO View Post
    If it were approaching zero, it would be approaching infinity. It was zero...and x/0 == undef.
    Touche! (or, in the immortal words of Buzz Lightyear, "To inifinity and beyond!")

    Definitely getting punchy...
    Last edited by HESmith; 02-20-2013, 01:16 PM. Reason: typo

    Leave a comment:


  • ECO
    replied
    Originally posted by HESmith View Post
    Since data = 0, don't you mean infinite?
    If it were approaching zero, it would be approaching infinity. It was zero...and x/0 == undef.

    Leave a comment:


  • scbaker
    replied
    Originally posted by ECO View Post
    Overall my conversation was pretty disappointing as far as new info (although the gentleman I talked to resisted a pretty good grilling from me and a 454/Roche product manager).

    Points from my conversation that I can remember...
    • GridION == "4-5x more sensors" than the MinION (no comparative runtime specs).
    • GridION can sip from a 96 well plate full of samples (this was demo'd as a flat bottom black microtiter plate with no septa...hmmm).
    • Acquisition speed == "bases per second".
    • The above picture suggesting sample prep is just a "10bp overhang" is not correct and not confirmed
    • The prototypes of the MinION ended up much larger than shown at AGBT due to heat issues (had to "add fans").
    • They will not confirm which gating protein (exo or phi29) or which pore (mspA or a-HL), but that the gate protein is not covalently coupled to the pore. (which strongly suggests to me that it is phi29...the exo-release method would undoubtedly need exquisite positioning for efficient detection)
    • According to the CEO, the DNA input concentration is "nanomolar". This makes sense to be given the binding kinetics of phi29 to it's binding site...but translates into high concentrations for a single molecule platform. When questioned about the possibility of doing very dilute samples where all the molecules matter, he suggested some method of on-chip enrichment/concentration (?????).
    • They are cagey about data release because they want to (my paraphasing) "release data when it's ready"

    Anyway, that's all I got. The ratio of ([booth girls] + [Apple-esque brushed aluminum] + [blinky lights]) to (data) was undefined...so that seems to speak for itself.
    Thanks for summarizing for those of us who couldn't be at ASHG. All indications up to now have been for ONT using a highly modified version of a-HL, so I'd be surprised if the first version actually uses mspA. In terms of the exonuclease method, I'm pretty sure it's been abandoned (or put waaaay back on the backburner - hence the 'arbitration hearing' between ONT and Illumina).

    Leave a comment:


  • HESmith
    replied
    Originally posted by ECO View Post
    The ratio of ([booth girls] + [Apple-esque brushed aluminum] + [blinky lights]) to (data) was undefined...
    Since data = 0, don't you mean infinite?

    Leave a comment:


  • ECO
    replied
    Overall my conversation was pretty disappointing as far as new info (although the gentleman I talked to resisted a pretty good grilling from me and a 454/Roche product manager).

    Points from my conversation that I can remember...
    • GridION == "4-5x more sensors" than the MinION (no comparative runtime specs).
    • GridION can sip from a 96 well plate full of samples (this was demo'd as a flat bottom black microtiter plate with no septa...hmmm).
    • Acquisition speed == "bases per second".
    • The above picture suggesting sample prep is just a "10bp overhang" is not correct and not confirmed
    • The prototypes of the MinION ended up much larger than shown at AGBT due to heat issues (had to "add fans").
    • They will not confirm which gating protein (exo or phi29) or which pore (mspA or a-HL), but that the gate protein is not covalently coupled to the pore. (which strongly suggests to me that it is phi29...the exo-release method would undoubtedly need exquisite positioning for efficient detection)
    • According to the CEO, the DNA input concentration is "nanomolar". This makes sense to be given the binding kinetics of phi29 to it's binding site...but translates into high concentrations for a single molecule platform. When questioned about the possibility of doing very dilute samples where all the molecules matter, he suggested some method of on-chip enrichment/concentration (?????).
    • They are cagey about data release because they want to (my paraphasing) "release data when it's ready"

    Anyway, that's all I got. The ratio of ([booth girls] + [Apple-esque brushed aluminum] + [blinky lights]) to (data) was undefined...so that seems to speak for itself.

    Leave a comment:


  • Scoob
    replied
    GridIron

    Hmm. My XBox 360 looks a lot like that.....

    Leave a comment:


  • h2so4hurts
    replied
    Good article

    Chief Executive Officer Gordon Sanghera spent some time telling me how the technology could tag proteins or microRNA with specially-made DNA strands and be used for detection, but he wouldn’t say a word about the company’s timeline for DNA sequencing. Instead, he urged patience. “What we said at AGBT, we will make good.”
    I thought they said beta units would be out by September or October and they'd go on sale in December. It'd be a lot easier to be patient if they spared us the dog and pony show and just delivered.

    Leave a comment:


  • Nanoporous
    replied
    "wait for Nanopore’s sequencing data has been like waiting for Godot"

    Leave a comment:


  • h2so4hurts
    replied
    Anyone can put a plexiglass top on a microATX case full of wires...Mike's post reflects my own opinions, I HOPE this isn't all just marketing. But it's really hard to not yell bull**** if they're not going to release data or show these things actually *doing* something.

    Leave a comment:


  • GW_OK
    replied
    Good blog post toning down the hype over at mikethemadbiologist's page

    Leave a comment:


  • GW_OK
    replied
    Einstein Epigenomics Einstein Epigenomics ‏@EpgntxEinstein
    Oxford Nanopore powered up #ASHG2012

    Leave a comment:


  • GW_OK
    replied
    Yaniv erlich ‏@erlichya
    Based on oxford nanopore representative explanations, here is a technical annotation of the MinIon picture #ashg2012

    Leave a comment:

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