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Compositions and methods for sample preparation

a technology of composition and method, applied in the field of composition and method for disrupting cells, can solve problems such as damage to dna samples, and achieve the effect of less affinity for nucleic acids

Inactive Publication Date: 2016-12-29
IBIS BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The method significantly increases DNA recovery by reducing surface energy and adsorption, allowing for efficient isolation from various samples, including those with low concentrations, and improving diagnostic capabilities.

Problems solved by technology

These processes are inherently laborious, may result in damaged DNA samples and are generally not amenable to automation.

Method used

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  • Compositions and methods for sample preparation
  • Compositions and methods for sample preparation

Examples

Experimental program
Comparison scheme
Effect test

example 1

Basic Reaction

[0032]To an existing tube of beads used for cell lysis (e.g. a tube containing ZY beads) add perfluorooctyltriethoxysilane. In one embodiment, to 900 mg of beads, 100 microliters of perfluorooctyltriethoxysilane is added. Agitate components. Allow reaction to proceed for 30 minutes to 1 hour at room temperature. Use beads without further purification.

example 2

Purification

[0033]To an existing tube of beads used for cell lysis (e.g. a tube containing ZY beads) add perfluorooctyltriethoxysilane. In one embodiment, to 900 mg of beads, 100 microliters of perfluorooctyltriethoxysilane is added. Allow the reaction to proceed for 30 minutes to 1 hour at room temperature. Agitate components. Remove excess reagent and byproducts through heating with reduced pressure or lyophilization.

example 3

Increased Reaction Time

[0034]To an existing tube of beads used for cell lysis (e.g. a tube containing ZY beads) add perfluorooctyltriethoxysilane. In one embodiment, to 900 mg of beads, 100 microliters of perfluorooctyltriethoxysilane is added. Allow the reaction to proceed overnight at room temperature. Agitate components. Remove excess reagent and byproducts through heating with reduced pressure or lyophilization.

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Abstract

The invention relates generally to compositions and methods for disrupting cells (e.g., disrupting cellular and nuclear membranes). In particular, the invention provides modified solid surfaces (e.g., bead surfaces) and their use in disruption of cellular membranes (e.g., during cellular lysis procedures (e.g., for recovery of nucleic acid (e.g., DNA or RNA) from mechanical cell lysis)). Compositions and methods of the invention find use in a wide range of applications including molecular biology and medical science.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application is a continuation of U.S. application Ser. No. 14 / 369,627 filed Jun. 27, 2014, which is a national phase application under 35 U.S.C. §371 of PCT International Application No. PCT / US2012 / 072073 filed Dec. 28, 2012, which claims priority to U.S. Provisional Application Ser. No. 61 / 581,504 filed Dec. 29, 2011, the entirety of each are incorporated by reference herein.FIELD OF THE INVENTION[0002]The invention relates generally to compositions and methods for disrupting cells (e.g., disrupting cellular and nuclear membranes). In particular, the invention provides modified solid surfaces (e.g., bead surfaces) and their use in disruption of cellular membranes (e.g., during cellular lysis procedures (e.g., for recovery of nucleic acid (e.g., DNA or RNA) from mechanical cell lysis)). Compositions and methods of the invention find use in a wide range of applications including molecular biology and medical science.BACKGROUND ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C12N15/10
CPCC12N15/1006C12Q1/6806
Inventor CHIESL, THOMAS N.
Owner IBIS BIOSCI
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