Method for preparing nucleic acid probe library

A technology of nucleic acid probes and probes, which is applied in biochemical equipment and methods, microbial measurement/inspection, fermentation, etc., and can solve the problem of inability to analyze or discover unknown genes, probe libraries that have not been widely used, and disposable High purchase cost and other issues, to achieve the effect of strong anti-strike ability, reduce dependence, and convenient transportation

Inactive Publication Date: 2006-07-05
TSINGHUA UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The disadvantage of this probe preparation method is that the one-time purchase cost is high; in addition, this probe uses a clear sequence, which can only be used to analyze known genes, but cannot analyze or discover unknown genes; moreover, The probe library itself cannot be copied, once it is used up, it needs to be purchased again
This kind of probe library has not been widely used at present, and in the existing applications, the evaluation of its effect is also mixed.

Method used

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  • Method for preparing nucleic acid probe library
  • Method for preparing nucleic acid probe library
  • Method for preparing nucleic acid probe library

Examples

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Effect test

Embodiment 1

[0017] Example 1: Preparation of Human Gene Cloning Probe Library

[0018] 1. Human gene probe cloning

[0019] Three clones 175C1, 175D3, and 175A5 of different lengths were randomly selected from the human gene clone library of the Beijing National Engineering Research Center for Biochips (the uniform number of the National Engineering Research Center for biochips), and the lengths of the human gene clones contained in them were respectively 490bp, 750bp and 1000bp, basically covering all clones in length. These clones will serve as templates for profiling microarray probe preparation in two different formats. The first form is preserved engineering bacteria culture, and the second form is plasmid prepared by alkaline lysis method.

[0020] 2. Primers

[0021] Conventional primer: upstream primer 5'CTG CAA GGC GAT TAA GTT GGG TAA C 3';

[0022] Downstream primer 5'GTG AGC GGA TAA CAA TTT CAC ACA GGA AAC AGC 3';

[0023] Universal primer: upstream primer 5'TCA ...

Embodiment 2

[0038] Example 2. Preparation of clinical infectious disease pathogen clone probe library

[0039] 1. Cloning of pathogenic probes for clinical infectious diseases

[0040] Four clones pTP205, pHCV244, pHIV309 and pHBV538 with different lengths were selected from the clinical infectious disease pathogen clone library of Beijing National Engineering Research Center of Biochip, and the lengths of the clinical infectious disease pathogen clones contained in them were 205bp and 244bp respectively , 309bp and 538bp. These clones correspond to four different pathogens: TP (Treponema pallidum), HCV (Hepatitis C virus), HIV (AIDS virus) and HBV (Hepatitis B virus). These clones will be in the form of plasmids as templates for microarray probe preparation.

[0041] 2. Primers

[0042] Conventional primers: Treponema pallidum: upstream primer 5'AAC ACG ATC CGC TAC GAC TAC TAC 3',

[0043] Downstream primer 5'CCC TAT ACC CGT TCG CAA TCA AAG 3';

[0044] ...

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Abstract

The present invention discloses a method for preparing nucleic acid probe library. It uses the oligonucleotide whose 5' has general sequence which is regardless of cloned probe sequence, carrier sequence and gene squence of culture bacteria and 3' has probe amplification sequence as primer, and uses the culture bacteria containing probe clone or plasmid as template to make amplification and obtain first-grade probe library, the uses the general sequence as primer and uses first-grade probe library as template to make amplification and obtain second-grade probe library, and the live, uses the probe amplification sequence as primer and uses the above-mentioned probe library as template to make amplification so as to obtain the nucleotic acid probe library.

Description

technical field [0001] The invention relates to a method for preparing a microarray probe library in the field of biotechnology, in particular to a method for preparing a nucleic acid probe library. Background technique [0002] Since the concept of "biochip" was proposed in the 1990s, biochip technology has far surpassed many traditional biotechnologies with its integration, miniaturization, and ease of automation. The market potential has attracted widespread interest from all over the world, and it has developed and improved by leaps and bounds in just 10 years, and new technologies and methods based on biochips have emerged in an endless stream. At present, biochips have developed from the original microarray chips and gene chips to many complicated categories, including protein chips, antibody microarrays, neuron chips, tissue chips, capillary chips, and lab-on-a-chip. While the technology itself is constantly improving, the application fields of biochips are also cons...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12P19/34
Inventor 陶生策程京马雪梅
Owner TSINGHUA UNIV
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