SIMULTANEOUS EXTRACTION AND SEPARATION OF RNA AND DNA FROM SINGLE CELLS USING ELECTROPHORETIC TECHNIQUES

Title:

SIMULTANEOUS EXTRACTION AND SEPARATION OF RNA AND DNA FROM SINGLE CELLS USING ELECTROPHORETIC TECHNIQUES

Document Type and Number:

WIPO Patent Application WO/2015/106146

Kind Code:

A3

Abstract:

Devices and methods for preparing RNA and DNA from single cells are disclosed. In particular, the invention relates to a method of simultaneously extracting RNA and DNA from single cells and separating the nucleic acids electrophoretically. An electric field is used to lyse a single target cell, such that the plasma membrane is selectively disrupted without lysing the nuclear membrane. Cytoplasmic RNA is separated from the nucleus by performing isotachophoresis (ITP) in the presence of a sieving matrix that preferentially reduces the mobility of the nucleus. During ITP, the cytoplasmic RNA accumulates at an ITP interface between leading and trailing electrolytes and can later be extracted free of nuclear DNA. The method can be performed in a microfluidic device that fully automates all steps of the process.

Inventors:

SANTIAGO JUAN G (US)
SHINTAKU HIROFUMI (US)

Application Number:

PCT/US2015/010884

Publication Date:

April 07, 2016

Filing Date:

January 09, 2015

Export Citation:

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Assignee:

UNIV LELAND STANFORD JUNIOR (US)

International Classes:

B01D57/02

Foreign References:

US20100224494A1	2010-09-09
US20100143878A1	2010-06-10
EP0320937A2	1989-06-21
US20040180350A1	2004-09-16
EP2073004A1	2009-06-24

Other References: