Sequencing by synthesis using mechanical compression

A method of sequencing by synthesis using mechanical compression, the method comprising:

• sequencing a library of polynucleotide fragments by:
• sequentially moving each droplet of a series of fluidic droplets within a cartridge, wherein the cartridge comprises an air gap formed between a first sheet having a first surface that is hydrophobic and oleophobic and a second sheet having a second surface that is hydrophobic and oleophobic, wherein the first sheet and the second sheet are secured opposite each other at a predetermined spacing relative to each other with the air gap therebetween, the cartridge further comprising the library of polynucleotide fragments, wherein the library of polynucleotide fragments comprises a plurality of clusters of polynucleotides on the second sheet, wherein moving each droplet comprises moving one or more mechanical force applicators against the first sheet to elastically deform the first sheet and create a region of reduced spacing between the first and second sheets so that each droplet follows the region of reduced spacing over the plurality of clusters of polynucleotides, wherein the series of fluidic droplets includes:
• a nucleotide polymerase reaction mix droplet, a first wash buffer droplet, a dye/terminator cleavage mix droplet, and a second wash buffer droplet, and
• imaging the plurality of clusters of polynucleotides to detect addition or hybridization of a nucleotide to clusters of the plurality of clusters of polynucleotides;
• and
• repeating the steps of sequentially moving each droplet of the series of fluidic droplets and imaging the plurality of clusters to generate sequence data for the plurality of clusters of polynucleotides.

A method of sequencing by synthesis using mechanical compression, the method comprising:

• sequencing a library of polynucleotide fragments by:
• sequentially introducing a series of fluidic droplets into an air gap formed between:
• a first sheet having a first surface that is hydrophobic and oleophobic;
• and
• a second sheet having a second surface that is hydrophobic and oleophobic, wherein the first sheet and the second sheet are secured opposite at a predetermined spacing relative to each other with the air gap therebetween, the second sheet further comprising the library of polynucleotide fragments, wherein the library of polynucleotide fragments comprises a plurality of clusters of polynucleotides on the second sheet;
• sequentially moving each droplet of the series of fluidic droplets over the plurality of clusters of polynucleotides by moving one or more mechanical force applicators against the first sheet to elastically deform the first sheet and create a region of reduced spacing between the first and second sheets so that each droplet follows the region of reduced spacing over the plurality of clusters of polynucleotides, wherein the series of fluidic droplets includes:
• a nucleotide polymerase reaction mix droplet, a first wash buffer droplet, a dye/terminator cleavage mix droplet, and a second wash buffer droplet;
• imaging the plurality of clusters of polynucleotides to detect addition or hybridization of a nucleotide to clusters of the plurality of clusters of polynucleotides;
• and
• repeating the steps of sequentially introducing each droplet of the series of fluidic droplets, moving each droplet and imaging the plurality of clusters to generate sequence data for the plurality of clusters of polynucleotides.