Originally posted by TonyBrooks
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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.
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Originally posted by ECO View PostOverall 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.
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.
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Originally posted by ECO View PostIf it were approaching zero, it would be approaching infinity. It was zero...and x/0 == undef.
Definitely getting punchy...
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Originally posted by ECO View PostOverall 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:
-
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.
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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.”
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"wait for Nanopore’s sequencing data has been like waiting for Godot"
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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.
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Einstein Epigenomics Einstein Epigenomics @EpgntxEinstein
Oxford Nanopore powered up #ASHG2012
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Yaniv erlich @erlichya
Based on oxford nanopore representative explanations, here is a technical annotation of the MinIon picture #ashg2012
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