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Repurposing beads in sample cleanup

A bead, magnetic bead technology, applied in the field of nucleic acid capture, which can solve the problems of inaccessibility of therapeutics, limited automation of RNA and DNA analysis, and increased cost

Pending Publication Date: 2021-08-20
帝肯基因组学公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using two different sets of beads in this manner limits the automation of RNA and DNA analysis and greatly increases the costs associated with early disease detection and drug development
This leaves millions of people with treatable diseases without access to adequate treatment

Method used

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  • Repurposing beads in sample cleanup
  • Repurposing beads in sample cleanup

Examples

Experimental program
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Effect test

example 1

[0025] Example 1: Benchtop method.

[0026] RNA-Seq libraries were generated in which all purification steps were performed using a single set of oligo dT magnetic beads (GE Health Care, Ser-Mag oligo dT beads). 100 ng of total RNA derived from the K562 cell line was mixed with 35 μl of Oligo dT beads following the supplier's recommendation. After annealing, the beads were pulled to the side of the tube and the solution containing any unbound RNA was discarded. Beads were resuspended and washed as recommended by the supplier. The beads were collected and the wash buffer was discarded. Bound RNA was eluted in 20 μl of RNA Fragmentation Buffer (Tecan Group Ltd., Universal RNA-Seq) and incubated at 94° C. for 7 minutes as recommended. The beads were collected, and the fragmented RNA was transferred to a new tube.

[0027] The oligo dT beads were resuspended in 40 μl of reconstitution solution (bead binding buffer) containing 21% PEG 4000, 2.5M NaCl, 50 mM Tris, 0.1 mM EDTA an...

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Abstract

The application relates to repurposing beads in sample cleanup. The present invention provides methods for repurposing beads and other solid supports for separately capturing RNA and DNA without a loss of binding capacity by using a surfactant.

Description

technical field [0001] The present invention relates to methods for capturing nucleic acids. Background technique [0002] Methods for DNA and RNA identification and analysis have become commonplace. Nucleic acids must often be isolated or captured from complex mixtures of prepared nucleic acids prior to analysis. For example, isolating mRNA is an important step in the analysis of gene expression and gene regulation, and can be invaluable in early disease detection and drug development. [0003] Common methods for DNA and RNA capture use magnetic solid supports such as magnetic beads. These methods bind DNA or RNA to beads by using one or more ligands on the bead surface. A wide variety of ligands are available that have an affinity for a particular particle. For example, mRNA is often captured using beads coated with a ligand that has an affinity for the adenosine (poly(A)+ tail) homopolymer at one end of the mRNA molecule. One such bead for mRNA capture is coated with...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1013C12Q1/6806C12Q2523/308C12Q2521/107C12Q2531/113C12N15/1096C12Q1/6874
Inventor D·A·阿莫雷塞
Owner 帝肯基因组学公司
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