[DIYbio] Re: How does Helicos see single molecule fluorescence?

Helicos is the only system that can generate usable reads directly from RNA and it is vastly superior to any other current system for quantitative accuracy in applications like ChIP Seq and RNA Seq where read length and accuracy are less important than number of reads and lack of amplification.  Despite the technical advantages, commercial uptake has been extremely limited for a variety of reasons and this is not improving.  A quick look at the stock price will confirm that.


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On Saturday, September 1, 2012 7:06:29 AM UTC-4, hrusty wrote:
Hello, I have been a follower of Helicos for a few years now. My question is to your technical expertise. Do you see this platform competitive in the current market or is it a league of its own because tSMS and DRS and lack of amplification making a direct threat to the staus quo?

Thanks for your time

On Thursday, August 23, 2012 5:00:29 PM UTC+2, Nathan McCorkle wrote:
This video says they take sheared ssDNA, add polyA tail, hybridize
about 1 molecule per square micron on a surface coated with polyT,
then add polymerase and a single species of fluorescent nucleotide,
rinse, illuminate with laser light and scan the surface with a camera,
cleave fluorophore, then repeat with a different species of nucleotide
and more polymerase.

http://www.helicosbio.com/Portals/0/Videos/tSMS-How_It_Works.flv

I wonder if they're using a single pixel camera with a PMT backend?

I wonder if I could replicate with just 1 pixel (1 molecule only), I
could use this to provide feedback for closed-loop DNA synthesis
operations.

--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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