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Whole blood DNA quick extraction kit

A kit and automatic extraction technology, applied in the biological field, can solve the problems of chemical toxicity, time-consuming and time-consuming, etc., achieving a high degree of automation and reducing human errors and experimental errors.

Inactive Publication Date: 2017-08-15
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the traditional method of extracting nucleic acid with phenol chloroform can obtain better quality nucleic acid, the chemical substances used in it have non-negligible toxicity, and the whole extraction process takes a long time, wasting a lot of time, manpower and material resources
Furthermore, the now popular column extraction nucleic acid extraction reagents can also obtain good quality nucleic acids with negligible toxicity, but the time-consuming can also not be ignored
[0004] In addition to toxicity and time factors, the addition and discarding of various washing solutions are inevitable in the operation of extracting nucleic acids with these two reagents. This process is extremely easy to cause cross-contamination between samples
When the sample size is large, it is easy to make mistakes, resulting in low work efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, a kind of preparation method of whole blood DNA rapid extraction kit:

[0031] a. Configuration of the lysate: first add the guanidine isothiocyanate weighed in the volumetric flask, then add Tris-HCl with a concentration of 15mmol / L and a pH of 3.3; add sodium hydroxide with a concentration of 5mmol / L; After the guanidine thiocyanate is completely dissolved, add 5% NP-40 and mix well, then add sterile water to the required volume.

[0032] b. Configuration of washing solution 1: first add a small amount of sterile water into the volumetric flask, add Tris-HCl with a concentration of 60mmol / L and a pH of 6.0; add ethanol with a concentration of 45%; mix well, and then add sterile water to required volume.

[0033] c. Configuration of washing solution 2: first add a small amount of sterile water to the volumetric flask, and then add CaCl with a concentration of 5mmol / L 2 ; Add 70% ethanol; mix well, add sterile water to the required volume.

[0034] d. C...

Embodiment 2

[0038] Embodiment 2, the application of a kind of whole blood DNA rapid extraction kit

[0039] Take whole blood as the sample, invert the pre-packed deep-well plate and centrifuge briefly for 10 seconds, then carefully tear off the sealing film, and add 200µl whole blood sample and 50µl protease to the 1st and 7th column K. Then set the extraction program on the nucleic acid extraction instrument according to the table below, and then perform nucleic acid extraction. When the program reaches the third step, the operation will be paused. At this time, open the door of the instrument, add 200 μl of binding solution to the sample wells (column 1 and column 7) one by one, and then click "continue to run". After the extraction program is finished, take out the pre-packed deep-well plate, and the solutions in columns 6 and 12 are the nucleic acid solutions.

[0040]

Embodiment 3

[0041] Example 3, comparison of two kits for nucleic acid extraction from whole blood samples

[0042] Taking 10 tubes of whole blood as samples, each tube was divided into two parts on average, and the nucleic acid extraction was compared with the kit of the present invention and the QIAamp DNA Blood Mini Kit (50)-Catalog No. 51104 kit of QIAGEN Company.

[0043] Kit extraction according to the present invention: nucleic acid extraction is carried out as described in Example 2; kit extraction by QIAGEN Company: nucleic acid extraction is carried out strictly according to the instructions of the QIAamp DNA Blood Mini Kit (50)-article number 51104 kit.

[0044] The nucleic acid extracted by the kit of the present invention and the QIAamp DNA Blood Mini Kit (50) of QIAGEN Company was subjected to OD value detection to obtain the nucleic acid concentration, and the results are shown in Table 1. It can be seen from the results in the table that the whole blood extraction efficienc...

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Abstract

The invention discloses a whole blood DNA quick extraction kit which comprises a lysis solution, a proteinase K, a combined solution, a magnetic bead, a cleaning solution 1, a cleaning solution 2 and an eluent. The 96-pore pre-split deep hole plate mode is adopted and a nucleic acid automatic extraction instrument can be combined for automatically extracting nucleic acid, so that the whole blood DNA can be quickly and effectively extracted in abundance.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a whole blood DNA rapid extraction kit. The invention also relates to a method for extracting whole blood DNA by using the magnetic bead method. Background technique [0002] In the diagnosis of modern diseases, nucleic acid plays a very important role, and the role of DNA is more prominent, and whole blood is an important source of DNA that is easy to obtain. In the process of PCR molecular diagnosis, the total copy number and quality of DNA obtained will directly affect the results of subsequent testing experiments. Therefore, obtaining effective and reliable DNA is the key to PCR molecular diagnosis. [0003] Although the traditional phenol-chloroform nucleic acid extraction method can obtain better quality nucleic acid, the chemical substances used in it have non-negligible toxicity, and the whole extraction process takes a long time, wasting a lot of time, manpower and materi...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 刘淑园陈华云肖湘文丁渭张天海邓金萍曾烨黄爽
Owner GUANGZHOU HEAS BIOTECH CO LTD
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