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US 11,180,800 B2Granted
Sample preparation methods, systems and compositions
AssigneeKarius, Inc.
InventorsTimothy A. Blauwkamp, Rene Sit, Igor D. Vilfan
Life of this patent
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Abstract
The disclosure provides methods, compositions, systems, and kits for the concurrent detection and analysis of different structural and chemical forms of nucleic acids in a sample.
Claims36
A method of performing an amplification reaction on a first RNA and a first DNA, comprising:
- a) providing a sample comprising a mixture of said first DNA and said first RNA, wherein said first DNA is cell-free DNA and does not comprise a sequence complementary to said first RNA;
- b) in said sample comprising said mixture of said first DNA and said first RNA, tagging said first DNA with a first tag without using a transposase;
- c) in said sample comprising said mixture of said first DNA and said first RNA, tagging said first RNA with a second tag;
- d) performing an amplification or primer extension reaction on said first DNA with a polymerase that is selective for DNA templates;
- and
- e) synthesizing a complementary DNA (cDNA) strand from said first RNA with a reverse transcriptase.
3 more elements
A method for concurrent processing of different nucleic acid forms in a sample comprising:
- a) denaturing said different nucleic acid forms in said sample, wherein said different nucleic acid forms in said sample comprise an RNA molecule and a DNA molecule;
- b) attaching a first adapter to one end of said RNA molecule and attaching a second adapter to one end of said DNA molecule wherein said DNA molecule is a cell-free DNA, thereby generating an RNA molecule attached to a first adapter and a DNA molecule attached to a second adapter;
- c) hybridizing a primer to said first adapter and performing an extension reaction in order to amplify said RNA molecule attached to said first adapter to generate an RNA extension product and hybridizing a primer to said second adapter and performing an extension reaction in order to amplify said DNA molecule attached to said second adapter to generate a DNA extension product;
- d) attaching a third adapter comprising a priming element to said RNA extension product and attaching a fourth adapter comprising a priming element to said DNA extension product wherein said third adapter and said fourth adapter comprise identical or different sequences to produce said RNA extension product attached to said third adapter and said DNA extension product attached to said fourth adapter;
- and
- e) amplifying said RNA extension product attached to said third adapter and said DNA extension product attached to said fourth.
3 more elements
Dependencies are inferred from the claim text.