Methods and compositions for polynucleotide library production, immortalization and region of interest extraction
US9096952B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 20, 2011 |
| Grant date | Aug 4, 2015 |
| Priority date | — |
| Expiry date | Oct 11, 2033 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6806
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Aspects of the present invention are drawn to methods and compositions for the genetic analysis of regions of interest (ROI) from one or more starting polynucleotide sample(s). In certain aspects, adapter tagged polynucleotide fragments from a plurality of initial polynucleotide samples are pooled, circularized and amplified to produce an immortalized library. Multiple ROI's from this immortalized library are amplified (e.g., in independent iPCR reactions) to generate amplicons, and, in some embodiments, pooled to form a pooled ROI amplicon sample. In certain embodiments, the amplicons for each ROI amplicon in the pooled ROI amplicon sample are present at known molar or mass ratios. The pooled ROI amplicon sample can be analyzed/processed as desired, e.g., sequenced using next generation sequencing technology.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.