METHODS AND SYSTEMS FOR NUCLEIC ACID ANALYSIS
The present disclosure provides methods and processes for increasing the efficiency and accuracy of nucleic acid sequencing using techniques such as polymerase chain reaction (PCR). Methods and systems provided herein may facilitate performing reactions such as emulsion PCR (ePCR) on samples comprising nucleic acids and beads. The methods provided herein may provide a higher throughput compared to existing technologies.
1 - 19 . (canceled)
20 . A method, comprising:
(a) using a thermocycler to adjust a temperature of a sample within a well of a plate disposed in a plate receptacle, wherein the plate receptacle comprises a thermal block, wherein beads from the sample and a first reagent are disposed in the well of the plate disposed in the plate receptacle;
(b) applying a magnetic force to the sample and the first reagent in the well of the plate to retain the beads within the well of the plate; and
(c) removing the first reagent from the well of the plate.
21 . The method of claim 20 , wherein (c) comprises using pressure or vacuum to remove the first reagent from the well.
22 . The method of claim 20 , wherein the thermal block comprises a heating element comprising a fluid.
23 . The method of claim 20 , wherein the magnetic force draws a bead of the beads toward a wall of the well to retain the bead within the well.
24 . The method of claim 20 , further comprising enriching a bead of the beads within the wells.
25 . The method of claim 24 , wherein the bead is coupled to a single template nucleic acid molecule.
26 . The method of claim 24 , wherein the bead is coupled to amplified nucleic acid molecules of a single template nucleic acid molecule.
27 . The method of claim 20 , further comprising thermocycling the sample.
28 . The method of claim 20 , wherein the sample comprises the beads.
29 . The method of claim 20 , wherein the plate comprises at least 96 wells.
30 . The method of claim 20 , wherein the plate is a 96-well plate.
31 . The method of claim 20 , wherein a bead of the beads is coupled to a nucleic acid molecule.
32 . The method of claim 31 , further comprising amplifying the nucleic acid molecule coupled to the bead in an amplification reaction.
33 . The method of claim 32 , wherein the amplification reaction comprises reverse transcription, primer extension, polymerase chain reaction (PCR), ligase chain reaction, helicase-dependent amplification, asymmetric amplification, rolling circle amplification, multiple displacement amplification (MDA), or a combination thereof.
34 . The method of claim 33 , wherein the amplification reaction comprises the PCR.
35 . The method of claim 34 , wherein the PCR comprises real-time PCR, allele-specific PCR, assembly PCR, asymmetric PCR, digital PCR, emulsion PCR (ePCR), dial-out PCR, helicase-dependent PCR, nested PCR, hot start PCR, inverse PCR, methylation-specific PCR, miniprimer PCR, multiplex PCR, nested PCR, overlap-extension PCR, thermal asymmetric interlaced PCR, touchdown PCR), ligase chain reaction (LCR), or a combination thereof.
36 . The method of claim 35 , wherein the PCR comprises the ePCR.
37 . The method of claim 31 , further comprising sequencing the nucleic acid molecule.
38 . The method of claim 26 , wherein the single template nucleic acid molecule comprises deoxyribose nucleic acid (DNA).
39 . The method of claim 26 , wherein the single template nucleic acid molecule comprises ribose nucleic acid (RNA).