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US 12,649,916 B2Granted
Method for selecting polynucleotides based on the size in a globular form
AssigneeOxford Nanopore Technologies PLC
InventorDaniel George Fordham
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Abstract
The invention provides methods of selecting polynucleotides based on their size in globular form. The polynucleotides may be treated with a condensation medium. e.g. comprising polyethylene glycol. e.g. PEG8000, and salt, in order to adopt a globular form. The method may employing filtering in order to retain the larger nucleic acids and make the larger nucleic acids available for sequencing applications, such as nanopore sequencing. The invention also relates to kits for selecting polynucleotides.
Claims15
A method comprising:
- (i) providing a sample comprising a mixture of polynucleotides and contacting the sample with a condensing medium, the condensing medium comprising about 20 mg/ml to about 150 mg/ml of a hydrophilic polymer and having a salt concentration from 0.1 M to 1 M, such that the polynucleotides in the sample adopt a globular form;
- wherein contacting the sample with a condensing medium does not precipitate the polynucleotides in the sample;
- (ii) filtering the sample through a filter such that polynucleotides retained by the filter are selected polynucleotides and polynucleotides passing through the filter are unselected polynucleotides, wherein the selected polynucleotides are longer than unselected polynucleotides,
- thereby selecting polynucleotides based on the size of the polynucleotides in the globular form;
- (iii) processing the selected polynucleotides such that the selected polynucleotides adopt a non-globular form,
- wherein the processing comprises contacting the selected polynucleotides with an extending medium;
- and
- (iv) sequencing the selected polynucleotides with a read length of at least about 25000 base pairs.
5 more elements
Dependencies are inferred from the claim text.