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Enzyme reaction method for nucleic acid and composition for separating nucleic acid

An enzymatic reaction and nucleic acid technology, applied in the field of nucleic acid, concentrating biological substances and transferring them from high-concentration salt solution to low-concentration salt solution, which can solve the problem that it cannot be used to remove inhibitors, and achieve the effect of easy preparation

Inactive Publication Date: 2007-01-10
ROCHE DIAGNOSTICS GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, this method cannot be used to remove substantial inhibitors of specific enzyme reactions that may be present

Method used

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  • Enzyme reaction method for nucleic acid and composition for separating nucleic acid
  • Enzyme reaction method for nucleic acid and composition for separating nucleic acid
  • Enzyme reaction method for nucleic acid and composition for separating nucleic acid

Examples

Experimental program
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Effect test

example 1

[0070] Preparation of magnetic particles of the present invention

[0071] Six sols were used. The sol was prepared as follows:

[0072] Sol 1 (SiO 2 :B 2 o 3 =7:3):

[0073] Synthesis was carried out in a 250 ml round flask with constant agitation.

[0074] 86.6ml tetraethylorthosilicate

[0075] +7ml absolute non-denatured ethanol

[0076] +14.1ml 0.15M HCl

[0077] A two-phase mixture was produced. Stir at room temperature until single phase. Add +37.8ml trimethyl borate drop by drop

[0078] The sol was then left at 50°C for 2 hours. join in

[0079] +14.1ml 0.15M HCl

[0080] Sol 2 (SiO 2 :B 2 o 3 =4:1):

[0081] Synthesis was carried out in a 250 ml round flask with constant agitation.

[0082] 100.5ml tetraethylorthosilicate

[0083] +7ml absolute non-denatured ethanol

[0084] +16.3ml 0.15M HCl

[0085] A two-phase mixture was produced. Stir at room temperature until single phase. Add +25.6ml trimethyl borate drop by drop

[0086] The sol was th...

example 2

[0126] Preparation of GMP1, GMP2, GMP3 and GMP4

[0127] GMP1, GMP2, GMP3 and GMP4 are different batches of pigments prepared under the following conditions from sol 1 (example 1) by the process described in example 1:

[0128] parameters

example 3

[0130] Pretreatment of human whole blood PCR samples using magnetic glass particles

[0131] nucleic acid isolation

[0132] 3 parts of glass magnetic particles (GMP2-4), each 10 mg into a micro test tube. The exact sample weights are given in Table 1. Triplicate determinations were performed.

[0133]40 μl proteinase K (20 mg / ml, made by lyophilization) was pipetted into 200 μl thawed whole blood and mixed immediately. Next, 200 μl of binding buffer (6M guanidine-HCl, 10 mM trimethylaminomethane-HCl, 10 mM urea, 30% Triton X-100, pH 4.4) was added and mixed, followed by incubation at 70° C. for 10 minutes. 200 μl of isopropanol was added and the formulation was mixed for 10 seconds in a vortex mixer. The samples were left at room temperature for 20 minutes and then mixed again for 10 seconds. Magnetic separation This step was performed for at least 30 seconds in a magnetic particle separator from Boehringer Mannheim (ID# 1 641 794). Supernatants were removed and analyze...

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PUM

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Abstract

New prods. are (a) magnetic particles with an external glass surface that is nonporous or has pores with a dia. below 10 nm and (b) ferromagnetic particles with a glass surface.

Description

[0001] This application was filed on June 6, 1996, the application number is "03106636.4", and the invention title is a divisional application of the invention patent application entitled "Method for Isolating Nucleic Acid". technical field [0002] The present invention relates to magnetic particles having a glass surface, and to methods of purifying biological substances, particularly nucleic acids, using glass particles in the presence of chaotropic salts. The invention also relates to methods of isolating these biological substances and methods of concentrating and transferring biological substances from high-concentration saline solutions to low-concentration saline solutions. Background technique [0003] Many biological substances, especially nucleic acids, present particular difficulties in isolating them from their natural environment. On the one hand, they are often present in very low concentrations, and on the other hand, they are often present together with many...

Claims

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Application Information

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IPC IPC(8): C12P19/34C12Q1/68G01N33/552B01J20/28B03CB03C1/01C07HC07H1/08C07H21/00C12NC12N15/09C12N15/10C12N15/11C12QG01N33/553H01F1/00H01F1/11H01F1/36
CPCY10S435/814H01F1/0063C12Q1/6804C12Q1/6834C03C3/087H01F1/36B03C1/01C03C3/091C03C3/111C03C3/078C03C3/108C03C3/085C07H21/00C03C3/083C03C3/102C03C3/105B82Y25/00C12N15/1013Y10S428/90C03C3/089C12Q1/6806H01F1/112Y10T428/2996
Inventor J·克莱恩T·沃尔特H·哈尔蒂希C·列斯尼亚克M·梅恩伊格M·里德林H·施密特
Owner ROCHE DIAGNOSTICS GMBH
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