Methods of using microfluidic positional encoding devices

A method of DNA assembly, comprising:

• disposing a solid phase support column in a chamber of a microfluidics circuit;
• performing an enzymatic reaction in a channel of the microfluidics circuit to produce a reaction mixture, the channel fluidly coupled to the chamber;
• and
• flowing the reaction mixture over the solid phase support column in the chamber to capture assembled oligos.

A method for tethered DNA assembly, comprising:

• capturing one or more oligonucleotide fragments on one or more solid supports within a microfluidic device;
• flowing additional oligonucleotide fragments across the captured oligonucleotide fragments;
• providing assembly enzymes to generate assembled DNA strands;
• flowing an error correction enzyme across the assembled DNA strands to create cleaved DNA strands and whole DNA strands;
• and
• flowing amplification primers across the assembled DNA strands to create support-free double stranded DNA, wherein the amplification primers only match the whole DNA strands.

A method for creating a unique molecular identifier for a molecule, comprising:

• providing a support unit having a plurality of attachment sites;
• attaching a molecule-specific multi-base oligomer at each of the plurality of attachment sites, wherein the molecule-specific multi-base oligomer is different for all attachment sites on the support unit;
• attaching a target-specific multi-base oligomer at at least a subset of the plurality of attachment sites on the support unit, wherein the target-specific multi-base oligomer is specific to a corresponding target molecule;
• and
• attaching the corresponding target molecule at each attachment site having the target-specific multi-base oligomer.

A bead sorting system, comprising:

• a microfluidic device comprising:
• a sorting junction configured to distribute a plurality of beads among a plurality of outlet channels;
• an inlet channel configured to route the plurality of beads to the sorting junction;
• a first steering channel intersecting the sorting junction and configured to steer fluid flow through the junction towards a first direction;
• and
• a second steering channel intersecting the sorting junction and configured to steer fluid flow through the junction towards a second direction;
• and
• a camera configured to monitor detection zones within a region of interest of the microfluidic device, the detection zones comprising:
• an actuation trigger zone located on the inlet channel upstream of the sorting junction;
• and
• a plurality of confirmation zones, each confirmation zone located on one of the plurality of outlet channels downstream of the sorting junction.

A method of assembling a barcoded sequence in a microfluidic device, comprising:

• generating a unique molecular identifier, the generating comprising:
• providing a support unit having a plurality of attachment sites;
• attaching a molecule-specific barcode at each of the plurality of attachment sites, wherein the molecule-specific barcode is different for all attachment sites on the support unit;
• and
• attaching a target-specific barcode at at least a subset of the plurality of attachment sites on the support unit, wherein the target-specific barcode is specific to a corresponding target molecule;
• and
• linking the unique molecular identifier to the corresponding target molecule via the target-specific barcode to create a barcoded strand of the corresponding target molecule.