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Quick extraction kit for blood sampling card

A kit and blood collection card technology, which is applied in the field of rapid extraction kits for blood collection cards, can solve problems such as unstable extraction efficiency, and achieve the effects of reduced manual operation time, simple operation, and uniform components

Inactive Publication Date: 2017-09-26
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the efficiency of proteinase K is not low, it is also a protein itself, and its performance is easily affected by factors such as the pH value and temperature of the external environment, resulting in unstable extraction efficiency, which in turn leads to differences between the same batch.

Method used

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  • Quick extraction kit for blood sampling card
  • Quick extraction kit for blood sampling card
  • Quick extraction kit for blood sampling card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: A kind of preparation method of quick extraction kit of blood collection card:

[0031] a. Preparation of blood slice digestion solution: first add Tris-HCl (pH 4.4) with a concentration of 1mol / L to the volumetric flask; after the Tris-HCl is mixed evenly, add EDTA (pH 8.0 ); after mixing EDTA, add NaCl with a concentration of 0.1mol / L; after mixing NaCl, add SDS with a concentration of 20g / L, and add sterile water to the required volume.

[0032] b. The selected concentration of tris(2-carboxyethyl)phosphine is 0.5 mol / L.

[0033] c. Configuration of the lysate: first add the guanidine isothiocyanate taken by weighing in the volumetric flask, add the Tris-HCl with a concentration of 12mmol / L and a pH of 3.2; add the sodium hydroxide with a concentration of 4.5mmol / L; After the guanidine isothiocyanate is completely dissolved, add NP-40 with a concentration of 6% and mix well; after the NP-40 is mixed well, add SDS with a concentration of 2% and mix well...

Embodiment 2

[0041] Embodiment 2: A kind of preparation method of blood collection card rapid extraction kit:

[0042] a. Configuration of blood slice digestion solution: first add Tris-HCl with a concentration of 3mol / L (pH 3.2) into the volumetric flask; after the Tris-HCl is mixed, add EDTA with a concentration of 0.8mol / L (pH10.0 ); after the EDTA is mixed, add NaCl with a concentration of 0.3mol / L; after the NaCl is mixed, add SDS with a concentration of 40g / L, and add sterile water to the required volume.

[0043] b. The selected concentration of tris(2-carboxyethyl)phosphine is 0.8 mol / L.

[0044] c. Configuration of the lysate: first add the guanidine isothiocyanate weighed in the volumetric flask, add Tris-HCl with a concentration of 14mmol / L and a pH of 2.8; add sodium hydroxide with a concentration of 2mmol / L; After the guanidine thiocyanate is completely dissolved, add NP-40 with a concentration of 10% and mix well; after the NP-40 is mixed well, add SDS with a concentration o...

Embodiment 3

[0052] Example 3: Application of a blood collection card rapid extraction kit

[0053] Take the blood collection card as the sample, first adjust the dry bath to 56°C, then cut the entire blood spot on the blood collection card into pieces, and transfer the pieces to 1.5ml EP tubes. Then add 400 μl of blood slice digestion solution and 50 μl of tris(2-carboxyethyl)phosphine, shake and mix well, place in a warm bath at 56°C for 1 hour, and mix well every 15 minutes. Finally, it was centrifuged at 12000 rpm for 1 min, and 200 μl of supernatant was extracted as a test sample.

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Abstract

The invention discloses a quick extraction kit for a blood sampling card. The quick extraction kit is prepared from the following components: blood film digestive juice, lysate, tris(2-carboxyethyl)phosphine, a binding solution, magnetic beads, a washing solution 1, a washing solution 2, a washing solution 3 and eluant. In order to increase the nucleic acid extraction efficiency of the kit, the tris(2-carboxyethyl)phosphine is used for replacing protease k, and polyethylene glycol (PEG)-6000 is added into the eluant. Furthermore, a 96-hole pre-subpackaged deep-well plate mode is used by the kit, and the kit can be matched with an automatic nucleic acid extractor-smart 32 of DA AN Gene Co. Ltd. of Sun Yat-sen University for use so as to realize automatic nucleic acid extraction; after the quick extraction kit is adopted, a great deal of deoxyribonucleic acid (DNA) of the blood sampling card can be rapidly and effectively extracted.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid extraction kit for a blood collection card. The invention also relates to a method for extracting the blood collection card 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. Blood collection card is an important means for preserving blood DNA. The stability of the blood collection card is high, which is consistent with the PCR result of the directly extracted sample. And the blood collection card can store blood samples under dry conditions and prevent the growth of bacteria and fungi, which is convenient for collection, transportation and archiving. In addition, 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, the key to ...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
CPCC12N15/1013C12Q1/6806C12Q2523/308
Inventor 陈华云肖湘文刘淑园刘孝礼潘培伟赵丽
Owner GUANGZHOU HEAS BIOTECH CO LTD
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