Nucleic acid capture, concentration, and purification
An example of a kit includes a flow cell assembly. The flow cell assembly includes a reaction chamber, a temperature controlled flow channel in selective fluid communication with an inlet of the reaction chamber, and a filter positioned in the temperature controlled flow channel. The reaction chamber includes depressions separated by interstitial regions and capture primers attached within each of the depressions. The filter is i) to block concentrated biological sample-polymer complexes generated in the temperature controlled flow channel at a first temperature, and ii) to allow passage of concentrated biological sample and polymer released from the complexes in the temperature controlled flow channel at a second temperature.
1. A kit, comprising:
a flow cell assembly including:
a reaction chamber having:
depressions separated by interstitial regions; and
capture primers attached within each of the depressions;
a temperature controlled flow channel in selective fluid communication with an inlet of the reaction chamber;
a filter positioned in the temperature controlled flow channel, the filter i) to block concentrated deoxyribonucleic acid (DNA)-methyl cellulose complexes generated in the temperature controlled flow channel at a first temperature, and ii) to allow passage of concentrated DNA and methyl cellulose released from the complexes in the temperature controlled flow channel at a second temperature;
a bypass line in fluid communication with an inlet of the temperature controlled flow channel and with an outlet of the temperature controlled flow channel;
a first bypass valve to control flow of a sample fluid to the inlet of the temperature controlled flow channel; and
a second bypass valve to control flow of the concentrated DNA and the methyl cellulose to the reaction chamber.
2. The kit as defined in claim 1 , further comprising a sample fluid including:
an aqueous carrier;
a DNA sample;
methyl cellulose;
a polymer that is chemically inert to DNA hybridization; and
a salt.
3. The kit as defined in claim 2 , wherein the polymer that is chemically inert to DNA hybridization is selected from the group consisting of polyethylene glycol having a weight average molecular weight ranging from about 500 to less than about 200,000, polyvinyl pyrrolidone, polyvinyl alcohol, and combinations thereof.
4. The kit as defined in claim 2 , wherein:
the DNA sample is present in the sample fluid at a first molar concentration ranging from about 1 pM to about 1 mM;
the methyl cellulose is present in the sample fluid in an amount ranging from about 0.5 wt % to about 20 wt % based on a total weight of the sample fluid;
the polymer that is chemically inert to DNA hybridization is present in the sample fluid in an amount ranging from greater than 0 wt % to about 20 wt % based on the total weight of the sample fluid; and
the salt is present in the sample fluid at a second molar concentration ranging from greater than 0 M to about 2 M.
5. The kit as defined in claim 2 , wherein:
the DNA sample is present in the sample fluid at a first molar concentration ranging from about 1 pM to about 1 mM;
the methyl cellulose is present in the sample fluid in an amount ranging from about 1 wt % to about 5 wt % based on a total weight of the sample fluid;
the polymer that is chemically inert to DNA hybridization is present in the sample fluid in an amount ranging from about 5 wt % to about 16 wt % based on the total weight of the sample fluid; and
the salt is present in the sample fluid at a second molar concentration ranging from 0.4 M to about 2 M.
6. The kit as defined in claim 5 , wherein the polymer that is chemically inert to DNA hybridization is polyethylene glycol having a weight average molecular weight ranging from about 500 g/mol to about 1000 g/mol.
7. The kit as defined in claim 5 , wherein the salt is sodium chloride.
8. The kit as defined in claim 2 , wherein the salt is sodium chloride; sodium bromide; sodium iodide; salts containing potassium, calcium, magnesium, or ammonium cations; and salts containing carbonate, sulfate, phosphate, or nitrate anions.
9. The kit as defined in claim 2 , wherein the DNA sample includes cell-free DNA, library DNA, whole genome amplified DNA, or combinations thereof.
10. The kit as defined in claim 2 , wherein the DNA sample includes a plurality of differently sized DNA inserts including small DNA inserts having from about 100 bases or base pairs to about 1,000 bases or base pairs and large DNA inserts having more than 1,000 bases or base pairs.
11. The kit as defined in claim 2 , wherein the aqueous carrier includes water, a salt solution, or a buffer solution including a tris(hydroxymethyl)aminomethane hydrochloride (TRIS-HCl) buffer, a tris(hydroxymethyl)aminomethane (TRIS) buffer, or a saline sodium citrate (SSC) buffer.
12. The kit as defined in claim 1 , wherein at least one surface of the temperature controlled flow channel includes a heating plate.
13. The kit as defined in claim 1 , wherein a number of the depressions in a predefined area ranges from about 100 per mm 2 to about 50 million per mm 2 with each depression having an average pitch ranging from about 50 nm to about 100 μm and an average volume ranging from about 1×10 −3 μm 3 to about 100 μm 3 .
14. The kit as defined in claim 1 , wherein the reaction chamber further has a polymeric hydrogel present within each of the depressions, and wherein the polymeric hydrogel is an acrylamide copolymer.
15. The kit as defined in claim 14 , wherein the capture primers are attached to the polymeric hydrogel within each of the depressions.
16. The kit as defined in claim 15 , wherein the polymeric hydrogel and the capture primers are not present on the interstitial regions.