Use of homologous recombinase to improve efficiency and sensitivity of single cell assays
AssigneeBio-Rad Laboratories, Inc.
InventorsRahul RATTAN, Marta GONZALEZ-PLASKY
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Abstract
Methods of decreasing bead aggregation and improving specificity of nucleic acid hybridization using non-specific nucleic acid binding proteins is described.
Claims20
A method of introducing oligonucleotide-linked beads into partitions, the method comprising,
- providing a bulk solution comprising (i) a plurality of the oligonucleotide-linked beads and (ii) an amount of a non-sequence specific nucleic acid binding protein sufficient to reduce aggregation by the beads;
- and
- introducing the beads into the partitions such that the number of beads in the partitions averages less than 5 beads per partition.
A solution comprising (i) a plurality of the oligonucleotide-linked beads and (ii) an amount of a non-sequence specific nucleic acid binding protein sufficient to reduce aggregation by the beads.
A method of introducing oligonucleotide-linked beads into partitions, the method comprising,
- providing a plurality of partitions, wherein the partitions comprise (i) an oligonucleotide-linked bead and (ii) a non-sequence specific nucleic acid binding protein, and (iii) a target nucleic acid;
- and
- hybridizing the oligonucleotide to the target nucleic acid in the presence of the non-sequence specific nucleic acid binding protein.
A method of detecting a nucleic acid in a microfluidic channel, the method comprising,
- flowing a nucleic acid sample in a microfluidic channel passed an immobilized oligonucleotide-linked bead in the presence of a non-sequence specific nucleic acid binding protein under conditions sufficient to allow for specific hybridization of the oligonucleotide to a target nucleic acid in the nucleic acid sample;
- and
- detecting hybridization of the target nucleic acid to the oligonucleotide.
Dependencies are inferred from the claim text.