Whole blood genome DNA extraction kit

A genome and kit technology, applied in the field of whole blood genomic DNA extraction kits, can solve the problems of inaccurate detection results, reduced sample DNA recovery efficiency, nucleic acid recovery efficiency, etc., and achieve the effect of ensuring accuracy

Inactive Publication Date: 2020-11-27
常州市武进人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The magnetic beads used in the extraction of nucleic acid by the magnetic bead method are coated with various active groups on the surface. Studies have shown that the ionic strength, pH and other conditions in the lysate will directly affect the activity of these functional groups, and then affect the amount of nucleic acid adsorbed by the magnetic beads.
Studies have shown that when the dilution of guanidine salt lysate is reduced below 60%, the efficiency of sample DNA recovery is sign...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] In Example 1, a kit for extracting genomic DNA from whole blood, including lysate, regulating beads, nano-magnetic beads, washing solution A, washing solution B and eluent;

[0031] The lysate comprises guanidine isothiocyanate, sodium N-lauroyl sarcosinate, guanidine hydrochloride, proteinase k and Tween20;

[0032] The regulating beads comprise a semipermeable membrane balloon, and sodium chloride is placed inside the semipermeable membrane balloon;

[0033] The nano-magnetic beads are ferric oxide coated with silicon oxide;

[0034] The washing liquid A comprises cetyltrimethylammonium bromide, Tris-HCl, guanidine isothiocyanate, dehydrated alcohol and FMES;

[0035] The washing solution B comprises Tris-HCl and absolute ethanol;

[0036] The eluent includes Tris-HCl and disodium EDTA.

[0037] Further, the pH value of the lysate is 5.0-7.5; the concentration of guanidine isothiocyanate in the lysate is 1 mM, the concentration of N-lauroyl sarcosinate is 0.5%, the...

Embodiment 2

[0051] In embodiment 2, a whole blood genomic DNA extraction kit, including lysate, adjustment beads, nano-magnetic beads, washing solution A, washing solution B and eluent;

[0052] The lysate comprises guanidine isothiocyanate, sodium N-lauroyl sarcosinate, guanidine hydrochloride, proteinase k and Tween20;

[0053] The regulating beads comprise a semipermeable membrane balloon, and sodium chloride is placed inside the semipermeable membrane balloon;

[0054] The nano-magnetic beads are ferric oxide coated with silicon oxide;

[0055] The washing liquid A comprises cetyltrimethylammonium bromide, Tris-HCl, guanidine isothiocyanate, dehydrated alcohol and FMES;

[0056] The washing solution B comprises Tris-HCl and absolute ethanol;

[0057] The eluent includes Tris-HCl and disodium EDTA.

[0058] Further, the pH value of the lysate is 5.0-7.5; the concentration of guanidine isothiocyanate in the lysate is 3mM, the concentration of N-lauroyl sarcosinate is 1%, the concentr...

Embodiment 3

[0072] In embodiment 3, a whole blood genomic DNA extraction kit includes lysate, adjustment beads, nano-magnetic beads, washing solution A, washing solution B and eluent;

[0073] The lysate comprises guanidine isothiocyanate, sodium N-lauroyl sarcosinate, guanidine hydrochloride, proteinase k and Tween20;

[0074] The regulating beads comprise a semipermeable membrane balloon, and sodium chloride is placed inside the semipermeable membrane balloon;

[0075] The nano-magnetic beads are ferric oxide coated with silicon oxide;

[0076] The washing liquid A comprises cetyltrimethylammonium bromide, Tris-HCl, guanidine isothiocyanate, dehydrated alcohol and FMES;

[0077] The washing solution B comprises Tris-HCl and absolute ethanol;

[0078] The eluent includes Tris-HCl and disodium EDTA.

[0079] Further, the pH value of the lysate is 5.0-7.5; the concentration of guanidine isothiocyanate in the lysate is 2mM, the concentration of N-lauroyl sarcosinate is 0.5%, the concentr...

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Abstract

The invention relates to the technical field of a DNA extraction kit, in particular to a whole blood genome DNA extraction kit. The whole blood genome DNA extraction kit comprises lysate, an adjustingbead, a nano magnetic bead, a washing solution A, a washing solution B and an eluent; the lysate contains guanidine thiocyanate, sodium N-lauroyl sarcosinate, guanidine hydrochloride, proteinase k and Tween 20; the adjusting bead contains a semipermeable membrane balloon, and sodium chloride is put in the semipermeable membrane balloon; the nano magnetic bead is ferroferric oxide wrapping siliconoxide; the washing solution A contains hexadecyl trimethyl ammonium bromide, Tris-HCl, guanidine thiocyanate, absolute ethyl alcohol and FMES; the washing solution B contains Tris-HCl and absolute ethyl alcohol; the eluent contains Tris-HCl and disodium EDTA. When the kit is in use, the adjusting bead absorbs moisture produced in a fragmentation process and adjusts concentration of the lysate, the recovery efficiency of nucleic acid cannot be adversely affected, and accuracy of a detection result is guaranteed.

Description

technical field [0001] The invention relates to the technical field of DNA extraction kits, in particular to a whole blood genome DNA extraction kit. Background technique [0002] Nucleic acid is the carrier of genetic information, the most important biological information molecule, and the main object of molecular biology research. Therefore, nucleic acid extraction is the most important and basic operation in molecular biology experimental techniques. Nucleic acid extraction refers to the process of separating the nucleic acid from the carrier by physical and chemical methods. At present, most medical institutions and scientific research fields in China need to extract nucleic acid from a large number of blood samples. Therefore, in order to ensure that high-concentration nucleic acids are extracted, new technologies are constantly being developed. [0003] Nucleic acid extraction methods are various, including traditional phenol-chloroform method, salting-out method, me...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2527/137
Inventor 陈文亚毛伦林季莉莉马爱金刘志清王利惠张金王佳佳黄婷婷
Owner 常州市武进人民医院
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