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DNA extracting method needing no tube transferring

An extraction method and nozzle technology, which is applied in the field of DNA extraction without tube transfer, can solve the problems of cross-contamination and DNA loss of different samples, and achieve the effect of avoiding cross-contamination, avoiding DNA loss, and simplifying the operation steps of DNA extraction

Inactive Publication Date: 2017-02-22
ANHUI SENPENG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above DNA extraction methods all need to transfer the solution containing DNA in different containers. Since the solution attached to the pipette tip, centrifuge tube wall, etc. contains DNA, this will inevitably cause the loss of part of the DNA. In addition, the transfer operation also requires There is a certain risk of cross-contamination between different samples; the invention relates to a DNA extraction method and reagents without tube transfer, which can avoid tube transfer operations during the DNA extraction process

Method used

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  • DNA extracting method needing no tube transferring

Examples

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Embodiment 1

[0033] Example 1 For the extraction of a total amount of 500 pg DNA micro sample

[0034] 1) Reagent preparation

[0035] a. Lysis solution: its components are 5 mol / l guanidine hydrochloride, 2% sodium lauroyl sarcosinate (m / v), 5% Tween 20, 100 mmol / l Tris-HCl, 100 mmol / l sodium citrate, pH 7;

[0036] b. Rinsing solution: its component is 80% ethanol;

[0037] c. Eluent: its components are 10 mmol / l Tris-HCl, 0.1 mmol / l EDTA-2Na, pH 8.0;

[0038] d. A set of combined tubes.

[0039] 2) DNA extraction

[0040] a. Take a sterile, DNA-free cotton swab, drop 0.5 μl Identifiler Plus PCR Kit 9947A positive control DNA (1ng / μl) on the cotton swab, the kit is provided by Appplied Biosystems; break off the swab Put the sub-head into the combination tube lysis tube (1), add 400 μl lysate solution, 15 μl proteinase K, vortex to mix, and heat at 50°C for 10 minutes;

[0041] b. Centrifuge at 12000 rpm for 1 minute, discard the lysis tube (1), discard the liquid in the collection ...

Embodiment 2

[0052] Example 2 DNA Extraction for Fingerprint Exfoliated Cells

[0053] 1) Preparation and collection of fingerprint samples: press the index finger on the clean glass window for 5 seconds, and wipe the fingerprint with a wet cotton swab for DNA extraction.

[0054] 2) DNA extraction

[0055] a. Put the sample in the above 1) into the combination tube lysis tube (1), add 500 μl lysate solution, 25 μl proteinase K, vortex to mix, and heat at 100°C for 20 minutes;

[0056] b. Centrifuge at 12000 rpm for 2 minutes, discard the lysis tube (1), discard the liquid in the collection tube (3), put the adsorption tube (2) back into the collection tube (3), add 500 μl to the adsorption tube (2) to rinse solution, centrifuged at 12000 rpm for 1.0 min;

[0057] c. Discard the liquid in the collection tube (3), put the adsorption tube (2) back into the collection tube (3), add 500 μl rinse solution into the adsorption tube (2), and centrifuge at 12000 rpm for 1.0 minute;

[0058] d. D...

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Abstract

The invention belongs to the technical field of nucleic acid extraction and purification and particularly relates to a DNA extracting method needing no tube transferring. The DNA extracting method includes the steps of firstly, preparing reagents, to be more specific, preparing lysate, washing liquid and eluent, and preparing a combined tube; secondly, extracting DNA, to be more specific, placing a sample into the inner-layer splitting tube of the combined tube, adding the lysate and protease k, then performing splitting, centrifuging, allowing liquid in the inner-layer splitting tube to be combined by a nucleic acid absorption film when the liquid penetrates the nucleic acid absorption film in a middle-layer DNA absorption tube, and separating to obtain purified DNA; thirdly, performing PCR amplification and electrophoresis. The DNA extracting method has the advantages that the tube transferring is not needed, the DNA extracting steps are simplified, and detecting time is shortened; DNA loss and cross contamination risks possibly caused by the tube transferring are avoided.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction and purification, and in particular relates to a DNA extraction method without tube transfer. Background technique [0002] DNA is the carrier of genetic information. For eukaryotes, nuclear genomic DNA is located in the nucleus. To obtain purified DNA, it is first necessary to crack the cell membrane, nuclear membrane and other structures to release the DNA, and then remove cell debris, proteins, Lipids, sugars and other impurities to purify DNA; if DNA needs to be extracted from swabs, feces, soil and other samples, it is also necessary to consider removing swabs, feces, soil and other impurities; currently, the available DNA extraction methods include Phenol-chloroform extraction method, magnetic bead method, silica gel membrane method, silica bead method, etc. [0003] The principle of the phenol-chloroform extraction method is to lyse the cells through the lysate, and use an ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/101C12Q2523/308C12Q2527/125C12Q2521/537
Inventor 刘香玲
Owner ANHUI SENPENG BIOTECHNOLOGY CO LTD
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