US 2019/0119721 A1 · SINGLE CELL RNA AND MUTATIONAL ANALYSIS PCR (SCRM-PCR): A METHOD FOR SIMULTANEOUS ANALYSIS OF DNA AND RNA AT THE SINGLE-CELL LEVEL

Abstract

A method of simultaneously analyzing RNA and DNA in a sample, the method comprising the step (a) contacting the sample with a reverse primer from a first primer pair directed to a target RNA region to effect reverse transcription of RNA into cDNA with a reverse transcriptase; (b) subsequently contacting the sample with (i) a forward primer from the first primer pair directed to a second cDNA region, (ii) a forward and a reverse primers from a second primer pair targeted to a DNA region, and (ii) a DNA polymerase to simultaneously amplify the target cDNA and target DNA region; and (c) analyzing the amplified target cDNA region and/or amplified target DNA region. Also encompassed are uses of the method to analyze gene expression and mutations, kits comprising primers, enzymes, buffers.

Claims20

Dependent claims

-36. (canceled)

A method of simultaneously analyzing RNA and DNA in a sample, the method comprising the steps of:

(a) contacting the sample with a reverse primer from a first primer pair, the reverse primer from the first primer pair being directed to a target RNA region, and a reverse transcriptase to effect reverse transcription of the RNA into cDNA;
(b) subsequently contacting the sample with:
(i) a forward primer from the first primer pair, the forward primer from the first primer pair being directed to a target cDNA region,
(ii) a reverse primer and a forward primer from a second primer pair, the reverse primer and forward primer from the second primer pair being directed to a target DNA region, and
(iii) a DNA polymerase
to simultaneously amplify the target cDNA region and the target DNA region;
and
(c) analyzing the amplified target cDNA region and/or the amplified target DNA region.

5 more elements

Dependencies are inferred from the claim text.

Facts

Application

15/541,295Filed 21 Jan 2016

Priority

SG 10201500472R21 Jan 2015

Published

25 Apr 2019

Term ends

Not estimatedThe source records don’t support an estimate.

Classification8 codes

C12Q 1/686
Nucleic acid amplification reactions: polymerase chain reaction [PCR]
C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6876
Involving nucleic acids: nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
C12Q 2537/143
The purpose or use of: multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple…

Single cell rna and mutational analysis pcr (scrm-pcr): a method for simultaneous analysis of dna and rna at the single-cell level

Life of this patent

Single cell rna and mutational analysis pcr (scrm-pcr): a method for simultaneous analysis of dna and rna at the single-cell level

Life of this patent

Abstract

Claims20