Nucleic Acid Extraction from Heterogeneous Biological Materials
AssigneeExosome Diagnostics, Inc.
InventorsWayne Comper, Leileata M. Russo, Johan Karl Olov Skog
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Abstract
Methods for extracting high quality nucleic acids from a heterogenous collection of nucleic acid-containing materials from a biological sample are disclosed. The heterogenous collection of nucleic-acid containing materials may contain cells or microvesicles, or both. The extractions obtained by the methods described herein are characterized by high yield and high integrity, making the extracted nucleic acids useful for various applications in which high quality nucleic acid extractions are preferred, e.g., a diagnosis, prognosis, or therapy evaluation for a medical condition.
Claims29
A method of extracting nucleic acid from a biological sample, comprising the steps of:
- a) obtaining a biological sample;
- b) performing a sample pre-processing step on the biological sample to obtain a fraction comprising a heterogeneous collection of nucleic acid-containing materials, wherein the heterogeneous collection of nucleic acid-containing materials comprises cells or microvesicles, or both;
- c) performing an operation selected from
- i) an extraction enhancement operation;
- ii) an affinity exclusion operation;
- and
- iii) an extraction enhancement operation and an affinity exclusion operation;
- and
- d) extracting nucleic acid from the resulting materials.
2-3. (canceled)
A nucleic acid extraction from a heterogeneous collection of nucleic acid-containing materials taken from a biological sample, wherein the heterogeneous collection of nucleic acid-containing materials comprise cells or microvesicles, and wherein the quantitative ratio of 18S rRNA to 28S rRNA detectable in the extraction is within the range of approximately 1:1 to approximately 1:2;
- and is preferably approximately 1:2.
A nucleic acid extraction from a heterogeneous collection of nucleic acid-containing materials taken from a biological sample, wherein the heterogeneous collection of nucleic acid-containing materials comprise cells or microvesicles, and wherein the nucleic acid yield is greater than or equal to 50 pg/ml from 1 ml or 20 ml of biological sample.
(canceled)
A kit for extracting nucleic acid from a biological sample, comprising:
- a) an affinity agent;
- b) an extraction enhancement agent;
- and
- c) a lysis buffer.
A kit for extracting nucleic acid from a biological sample, comprising:
- a) an affinity agent;
- b) optionally, a lysis buffer or an extraction enhancement agent;
- and
- c) instructions for using the kit, located either in the package with the other kit components or in a different location accessible to a kit user, wherein the instructions comprise instructions for using the affinity agent in an affinity exclusion operation.
Dependencies are inferred from the claim text.