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Methods and devices for sequencing nucleic acids in smaller batches

  • US 9,145,589 B2
  • Filed: 08/19/2014
  • Issued: 09/29/2015
  • Est. Priority Date: 02/05/2007
  • Status: Active Grant
First Claim
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1. A method for carrying out steps of a nucleic acid sequencing protocol, comprising:

  • a) providing;

    i) a plurality of reservoirs, at least one of which comprising nucleic acid sequencing reagents, and at least one of which comprising wash buffer;

    ii) first nucleic acids in a flow path of a first flow cell and second nucleic acids in a flow path of a second flow cell, said first and second nucleic acids comprising templates to be sequenced, each of said flow paths associated with an inlet port and outlet port, each of said inlet ports and outlet ports associated with tubing, said flow cells mounted on iii) a moveable support, said support configured to move said flow cells to iv) a plurality of stations, at least some of which are reagent delivery stations and at least one comprising a wash station, each station in fluidic communication with at least one of said reservoirs via tubing, each station comprising station ports capable of being aligned with said flow cell inlet and outlet ports, each station port comprising an elastomeric seal;

    b) moving said moveable support under conditions such that said first flow cell is aligned with and connected to a reagent delivery station, such that said reagent delivery station ports are aligned with said flow cell inlet and outlet ports at interfaces sealed by said elastomeric seals, said seals defining an open area where fluids may flow, whereby said first flow cell is in fluid communication with at least a first reservoir of said plurality of reservoirs;

    c) introducing reagent via one of said reservoirs into said first flow cell under conditions such that said reagent contacts said first nucleic acids in said flow path of said first flow cell;

    d) disconnecting said first flow cell from said reagent delivery station associated with said first step;

    e) moving said moveable support under conditions such that said first flow cell is aligned with and connected to a wash station, such that said wash station ports are aligned with said flow cell inlet and outlet ports at interfaces sealed by said elastomeric seals, said seals defining an open area where fluids may flow, whereby said first flow cell is in fluid communication with a wash reservoir comprising wash buffer, said open area defined by said seals of said wash station port being greater than said open area defined by said seals of said reagent delivery station, so as to allow any reagents trapped in the sealing area to be washed away, thereby preventing contamination of flow cells with reagents from a previous step; and

    f) introducing wash buffer from said wash reservoir into said first flow cell.

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