Process for simultaneously carrying out RNA and DNA extraction from clinical samples by employing virus cracking liquid and precipitation process

A lysate and sample technology, applied in the direction of biochemical equipment and methods, microbial measurement/testing, sugar derivatives, etc., can solve the problem of RNA inappropriateness

Active Publication Date: 2005-07-06
SUZHOU SYM BIO LIFESCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are several problems with this method: first, this extraction method is only suitable for DNA, not for RNA; second, since nucleic acid cannot be seen by naked eyes, it is very easy to accidentally discard it during washing and other steps

Method used

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  • Process for simultaneously carrying out RNA and DNA extraction from clinical samples by employing virus cracking liquid and precipitation process
  • Process for simultaneously carrying out RNA and DNA extraction from clinical samples by employing virus cracking liquid and precipitation process
  • Process for simultaneously carrying out RNA and DNA extraction from clinical samples by employing virus cracking liquid and precipitation process

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0020] Simultaneous preparation and detection of virus samples containing HBV and HCV

[0021] 1. Collection of Serum and Plasma Samples

[0022] Collect whole blood samples into blood collection tubes without any anticoagulant, let stand at room temperature for 30 minutes, centrifuge at 4000rpm for 5 minutes in a desktop centrifuge, collect upper serum samples, mix well, let stand at room temperature for 30 minutes, and collect upper plasma samples.

[0023] 2. Sample Preparation

[0024] Add 200 μl of serum or plasma sample to 400 μl of virus lysate, shake and mix on a shaker, and heat in a 65°C water bath for 15 minutes. Take out, equilibrate the sample to room temperature, add 600 μl of isopropanol, centrifuge at 13000 rpm for 15 minutes at room temperature, discard the supernatant, collect the blue precipitate, wash with 75% ethanol, and place the obtained blue precipitate at 65°C Dry for 10 minutes, add 50 μl of buffer solution (10 mmole / L Tris.HCl pH6.5) to dissolve t...

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Abstract

The invention relates to a process for extracting RNA and DNA simultaneously. The process comprises the steps of: cracking the sample to be detected using virus cracking liquor; concentrating the nucleic acid (RNA and DNA) in said sample by precipitation; drying the precipitate and storing it at a temperature of -20íµ or directly performing nucleic acid amplification after dissolution. The process has a stable yield, and quality index control can be carried out for the entire preparation process.

Description

technical field [0001] The invention relates to a nucleic acid extraction technology, in particular to a method for simultaneously preparing DNA and RNA in clinical samples by using a virus lysate combined with a precipitation method. Background technique [0002] In the past ten years, the speed of development of molecular biology can be described as rapid progress, and genetic engineering is one of the most important ones, which has been widely used in clinical medicine, pharmacy, agriculture, forensic science, etc. important developmental significance. [0003] For the extraction of sample nucleic acid, the more classic method is to use organic solvent centrifugation. Extract with phenol and centrifuge with ethanol or isopropanol, then wash and dry. There are several problems with this method: first, this extraction method is only suitable for DNA, not for RNA; second, since nucleic acid cannot be seen with the naked eye, it is very easy to accidentally discard it durin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C12P19/34C12Q1/68
Inventor 黄道培杨国翠龚华斐
Owner SUZHOU SYM BIO LIFESCI CO LTD
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