Separating polynucleotide fragments
Provided is a method, including stretching a polynucleotide over a substrate including a plurality of equally spaced cleavage regions including a plurality of transposases, cleaving the polynucleotide with two or more of the plurality of transposases to form a plurality of polynucleotide fragments, and separating, within the plurality of polynucleotide fragments, a population of longer polynucleotide fragments from a population of shorter polynucleotide fragments. Also provided is a method including stretching a polynucleotide over a substrate including a plurality of equally spaced cleavage regions including a plurality of transposases, cleaving the polynucleotide with two or more of the plurality of transposases to form a plurality of polynucleotide fragments, and separating, within the plurality of polynucleotide fragments, a population of longer polynucleotide fragments from a population of shorter polynucleotide fragments.
1 . A method, comprising
extending an end of a polynucleotide through a pore of a membrane, wherein the polynucleotide is attached to a bead, the pore is narrower than the bead, and a side of the membrane opposite the bead comprises a nuclease wherein the nuclease comprises a transposase, and
cleaving the polynucleotide with the nuclease.
2 . The method of claim 1 , wherein the extending comprises electrophoresing.
3 . The method of claim 1 , wherein the polynucleotide is double-stranded.
4 . The method of claim 1 wherein the nuclease is attached to the membrane.
5 . The method of claim 1 , wherein the polynucleotide is single-stranded.
6 . The method of claim 1 , wherein the cleaving comprises forming a cleaved end and further comprising adding an adapter to the cleaved end.
7 . The method of claim 6 , wherein the polynucleotide is double-stranded and the cleaving comprises forming complementary ends and adding adapters to the complementary ends, wherein the adapters are not connected to each other.
8 . The method of claim 6 , wherein the polynucleotide is double-stranded and the cleaving comprises forming complementary ends and adding adapters to the complementary ends, wherein the adapters are connected to each other.
9 . The method of claim 8 , wherein the adapters comprise a hairpin loop.
10 . The method of claim 1 , further comprising releasing the polynucleotide from the bead.
11 . The method of claim 1 , further comprising synthesizing a copy of a strand of the polynucleotide.
12 . The method of claim 6 further comprising synthesizing a copy of a strand of the polynucleotide including the adapter.
13 . The method of claim 7 further comprising synthesizing a copy of a strand of the polynucleotide including the adapter.
14 . The method of claim 4 , wherein polynucleotide is double stranded and the transposase tagments the polynucleotide adding an adapter.
15 . The method of claim 14 further comprising synthesizing a copy of a strand of the polynucleotide including the adapter.
16 . The method of claim 14 further comprising releasing the polynucleotide from the bead.
17 . The method of claim 16 further comprising synthesizing a copy of a strand of the polynucleotide including the adapter.
18 . The method of claim 15 further comprising amplifying the copied strand of the polynucleotide.
19 . The method of claim 17 further comprising amplifying the copied strand of the polynucleotide.
20 . The method of claim 12 further comprising amplifying the copied strand of the polynucleotide.