Compositions and method for measuring and calibrating amplification bias in multiplexed PCR reactions
US9371558B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jan 9, 2015 |
| Grant date | Jun 21, 2016 |
| Priority date | — |
| Expiry date | Jan 9, 2035 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/166
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Compositions and methods are described for standardizing the DNA amplification efficiencies of a highly heterogeneous set of oligonucleotide primers as may typically be used to amplify a heterogeneous set of DNA templates that contains rearranged lymphoid cell DNA encoding T cell receptors (TCR) or immunoglobulins (IG). The presently disclosed embodiments are useful to overcome undesirable bias in the utilization of a subset of amplification primers, which leads to imprecision in multiplexed high throughput sequencing of amplification products to quantify unique TCR or Ig encoding genomes in a sample. Provided is a composition comprising a diverse plurality of template oligonucleotides in substantially equimolar amounts, for use as a calibration standard for amplification primer sets. Also provided are methods for identifying and correcting biased primer efficiency during amplification.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.