Anirban Chatterjee

US Patent:

20110313145, Dec 22, 2011

Filed:

Jun 10, 2011

Appl. No.:

13/157929

Inventors:

Andre Sharon - Newton MA, US
Anirban Chatterjee - Malden MA, US
Paul Mirsky - Jamaica Plain MA, US
Alexis Sauer-Budge - Lincoln MA, US

Assignee:

FRAUNHOFER USA, INC. - Plymouth MI
TRUSTEES OF BOSTON UNIVERSITY - Boston MA

International Classification:

C07H 1/08
B01D 35/00
C12M 1/24

US Classification:

536 2541, 4353061, 422535

Abstract:

Disclosed herein are methods for purification of RNA from a sample. The RNA can be total RNA or mRNA. The method involves preparing the sample in a solution of lysis buffer and depositing into a first end of a lysis straw such that the sample solution flows through the matrix of the lysis straw and is eluted from the opposite end of the lysis straw, and depositing the eluted material into a first end of a solid phase extraction (SPE) straw, such that the deposited solution flows through the matrix of the SPE straw towards the opposite end of the SPE straw, and eluting the RNA from the SPE straw by depositing a solution of elution buffer, into the first end of the SPE straw, such that the deposited solution flows through the matrix of the SPE straw and is eluted from the opposite end of the SPE straw, wherein purified RNA from the sample is present in the eluate of the SPE straw. When the RNA is total RNA, and the sample is a cell sample, and step b) requires adding a precipitating solution to the eluted material from step a), and depositing the solution into the first end of the SPE straw, wherein the straw comprises silica microspheres, such that the deposited solution flows through the matrix of the SPE straw toward the opposite end of the SPE straw. When the RNA is mRNA and step b) further comprises depositing the solution into the first end of the solid phase extraction (SPE) straw, wherein the straw comprises oligo-dT. Pressure (e.g., at least 10 psi pressure) and heat (e.g., about 60 C.) can be applied to the samples, and/or eluates and/or straws. Examples of lysis buffers, loading buffers and elution buffers are provided. Also disclosed are the specific straws and porous polymer monolith matrix components.