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Kit for separating RNA (ribonucleic acid) bound in RNA binding protein

A technology for binding proteins and kits, which is applied in the field of molecular biology and can solve the problems of many influencing factors, easy pollution and low yield.

Inactive Publication Date: 2013-08-07
CHINA PHARM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the methods currently used are different, and there are many influencing factors. It is easy to contaminate the sample, and the RNA is easily affected by the widespread RNase, which often leads to extremely low final yield and the quality of RNA cannot be guaranteed.

Method used

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  • Kit for separating RNA (ribonucleic acid) bound in RNA binding protein
  • Kit for separating RNA (ribonucleic acid) bound in RNA binding protein
  • Kit for separating RNA (ribonucleic acid) bound in RNA binding protein

Examples

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Embodiment 1

[0022] Isolation of mRNA bound by RNA-binding protein HuR in Raw264.7 cells after LPS stimulation

[0023] 1. Reagent source:

[0024] (1) Magnetic bead A is Dynabeads of Invitrogen Company;

[0025] (2) The magnetic bead G is Dynabeads of Invitrogen Company;

[0026] (3) Mouse IgG of the present invention is produced by Sigma Company;

[0027] (4) RNase inhibitor is for the production of RNase OUT for Invitrogen;

[0028] (5) Protease mixed inhibitor produced by Roche;

[0029] (6) KCl, MgCl 2 , Hepes, surfactant Nonidet P-40, dithiothreitol, 4-hydroxyethylpiperazineethanesulfonic acid are all produced by Sigma.

[0030] 2. Instrument source:

[0031] (1) Refrigerated centrifuge, Sigma;

[0032] (2) Microplate reader, iQ5Multi color Red-time PCR Deteetion System, all are Bio-Rad;

[0033] (3) Four-dimensional rotator, Qilin Bell;

[0034] (4) Magnetic stand, Invitrogen;

[0035] (5) Gene Quent Nucleic Acid Quantitative Instrument, GE Company;

[0036] 1. Preparatio...

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Abstract

The invention relates to the field of molecular biology, particularly a kit for separating RNA (ribonucleic acid) bound in an RNA binding protein. The kit provided by the invention is used for separating, extracting and purifying RNA bound in a specific RNA binding protein. The kit comprises a lysis solution, a buffer solution and magnetic beads. The reagents in the kit are proportionally improved and optimized, and thus, can simply and stably obtain the mRNA (messenger ribonucleic acid) bound in the specific RNA binding protein in cells at higher yield through antibody antigen reaction and magnetic field principle. The kit is used for further cDNA real-time quantification, micro-array chip detection and other subsequent researches.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for isolating RNA bound to RNA binding proteins. Background technique [0002] The regulation of gene expression plays an important role in the growth, differentiation and stress response of cells in organisms. However, in addition to regulating the transcription process through transcription factors, the post-transcriptional regulation of cells also plays an important role in physiological processes, including the specificity of mRNA transcription speed through RNA-binding proteins (RBPs). and quick regulation. At present, the research on the regulation mechanism of mRNA by RBPs and non-coding RNA has just begun. RNA immunoprecipitation (RNA immunoprecipitating, RIP) is a technique based on the antigen-antibody reaction to study the relationship between RNA transcripts that specifically bind to RNA binding proteins (RNAbindingproteins, RBPs) in vivo through co-immunoprec...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 张陆勇张爽孙丽新
Owner CHINA PHARM UNIV
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