Quantitative detection and analysis of molecules
AssigneeIllumina, Inc.
InventorRobert Meltzer
Life of this patent
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Abstract
The invention provides systems and methods for making sequencing libraries that are useful for quantitatively analyzing nucleic acids in a sample. Sample nucleic acids are randomly cleaved at, and PCR handled are attached to, a random cut site. The nucleic acid is amplified into a sequencing library in which a sequencing primer generates a sequence read from adjacent the random cut site. The sequence reads can be mapped to a reference, but they will also include a unique identifier sequence that comes from within the nucleic acid molecule being analyzed, i.e., an intrinsic molecular identifier (IMI). The IMI is unique for each molecule and can thus be used to deduplicate sequence reads originating from the same molecule.
Claims14
1-7. (canceled)
A method for generating a nucleic acid library, the method comprising:
- capturing RNA molecules from a single cell with capture oligos that include a first adapter;
- extending the capture oligos in the presence of template switching oligonucleotides (TSOs) to form first duplexes comprising the RNA molecules and cDNA;
- amplifying the cDNA to form second duplexes comprising the cDNA;
- and
- cleaving the second duplexes at, and attaching second adapters to, random cut sites to thereby form constructs that each include a label defined by intrinsic sequence of a cDNA segment adjacent the random cut site.
Dependencies are inferred from the claim text.