High-flux simple and convenient purification method by magnetic beads for sequencing product

A purification method and high-throughput technology, which are applied in the field of high-throughput, simple and rapid magnetic bead purification of sequencing products, and can solve the problems of high price and long purification time.

Inactive Publication Date: 2015-01-21
PEKING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, the commonly used purification methods in sequencing companies or laboratories are the alcohol / sodium acetate (alcohol / NaAc) method or the Centricon-100 purification column (PN: N930-2119) method. The alcohol / sodium acetate method takes a long time to purify, and takes about 120 minutes. Purification column method is time-consuming but expensive

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  • High-flux simple and convenient purification method by magnetic beads for sequencing product
  • High-flux simple and convenient purification method by magnetic beads for sequencing product

Examples

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Embodiment 1

[0026] Example 1: A high-throughput, simple and rapid method for purification of sequencing products using magnetic beads:

[0027] (1) Place 10 μL of sequencing PCR product in a 96-well plate;

[0028] (2) Add 10 μL of Croning BCB-300 magnetic beads to the 96-well plate, then add 41 μL of 85% alcohol, shake and mix;

[0029] (3), stand at room temperature for 3 minutes, then adsorb on the AM10027 magnetic stand of Ambion Company for 3 minutes to make the solution clear and then remove the supernatant;

[0030] (4), then add 100 μL of 85% alcohol and let it stand for 30 sec, then remove the supernatant;

[0031] (5), repeat step (4) once, then remove supernatant;

[0032] (6) Remove the 96-well plate from the AM10027 magnetic stand of Ambion, and place it on a 96-well heating block at 50°C for 30sec to 1min;

[0033] (7) Add 30 μL of sterilized deionized water to the 96-well plate, shake and mix, centrifuge for 10 sec, and let stand at room temperature for 3 min to comple...

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Abstract

The invention provides a high-flux simple and convenient purification method by magnetic beads for a sequencing product, belongs to genetic engineering, and relates to the technical field of purification method of DNA or RNA. The purification method comprises the following steps: placing a PCR product in a 96-mesh plate; adding magnetic beads in the 96-mesh plate; adding alcohol; oscillating for uniform mixing; leaving the materials to stand at a room temperature; conducting adsorption on a magnetic frame; clearing the solution; removing the alcohol; adding alcohol to stand; removing the alcohol; repeating the steps for one time; removing the alcohol; taking down the 96-mesh plate from the magnetic frame; placing the 96-mesh plate on a 50 DEG C 96-mesh heating block; adding sterilization deionized water in the 96-mesh plate; oscillating for uniform mixing; leaving the solution to stand at the room temperature after centrifugation; placing the 96-mesh plate on the magnetic frame to obtain the product purified by the magnetic beads. The purification method has the advantages that dyestuffs which do not react completely are removed more completely; the deion is economic, convenient and practical; dye peak can be removed effectively.

Description

technical field [0001] The invention belongs to the technical field of DNA or RNA purification methods involved in genetic engineering, and in particular relates to a high-throughput, simple and fast magnetic bead purification method for sequencing products. Background technique [0002] In the course of molecular biology experiments, plasmids or PCR products are usually sequenced routinely. How to purify the sequencing products has a great impact on the speed and quality of the sequencing results. Currently, the commonly used purification methods in sequencing companies or laboratories are the alcohol / sodium acetate (alcohol / NaAc) method or the Centricon-100 purification column (PN: N930-2119) method. The alcohol / sodium acetate method takes a long time to purify, and takes about 120 minutes. The purification column method, although time consuming, is expensive. Contents of the invention [0003] The purpose of the present invention is to disclose a high-throughput, simpl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 董志李桂澜
Owner PEKING UNIV
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