Nucleic acid isolation methods
AssigneeFluidigm Corporation
InventorsAndrew May, Alain Mir, Ramesh Ramakrishnan, Bernhard G. Zimmermann
Life of this patent
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Abstract
The present invention provides methods for selectively enriching a biological sample for short nucleic acids, such as fetal DNA in a maternal sample or apoptic DNA in a biological sample from a cancer patient and for subsequently analyzing the short nucleic acids for genotype, mutation, and/or aneuploidy.
Claims14
A method of enriching a sample for short nucleic acids of less than 300 nucleotides in length, the method comprising:
- contacting the sample with polyethylene glycol in the presence of a monovalent salt under conditions suitable for substantially precipitating nucleic acids of 300 or greater nucleotides in length, without substantially precipitating the short nucleic acids;
- and
- recovering the short nucleic acids.
A method of producing one or more target amplicons comprising one or more nucleotide tags from a sample, the method comprising:
- preamplifying the one or more target nucleic acids in the sample with a first, outer primer pair to produce one or more first target amplicon(s);
- and
- preamplifying the one or more target amplicon(s) with a second primer pair, wherein one or both primers of the second primer pair anneals to a primer binding site in between the primer binding sites of the outer primer pair, and
- wherein at least one of the first, outer primer pair and/or the second primer pair comprises a nucleotide tag.
A method of enriching a sample for short nucleic acids of less than 300 nucleotides in length, the method comprising:
- circularizing the sample nucleic acids under conditions that favor the circularization of short nucleic acids;
- and
- recovering the circularized nucleic acids.
A method for tagging long nucleic acids and removing long nucleic acids of greater than 200 nucleotides in length, the method comprising:
- contacting nucleic acids in the sample with a transposase in a reaction mixture under conditions suitable for fragmenting and tagging nucleic acids of greater than 200 nucleotides in length with transposon ends;
- and
- selecting and removing transposon-tagged nucleic acids from the reaction mixture by affinity selection.
Dependencies are inferred from the claim text.