Novel method and kit for extracting high-quality RNA (Ribose Nucleic Acid) from paraffin-embedded tissue
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A paraffin-embedded, kit technology, applied in recombinant DNA technology, DNA preparation and other directions, can solve the problems of small RNA fragments, difficult RNA extraction, easy degradation, etc., achieve long RNA length, save research funds, species information full effect
Inactive Publication Date: 2011-07-06
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
View PDF0 Cites 9 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0004] The purpose of the present invention is to provide a new method and kit that can simply and effectively extract high-quality RNA from conventional pathological paraffin tissues, so as to solve th
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0034] Example 1 Using the method of the present invention to extract RNA from multiple lung cancer tissues
[0035] The kit formula is the preferred formula:
[0036] ⑴Reagent A: The ingredient is a non-toxic, environmentally friendly and transparent dewaxing agent;
[0037] ⑵Reagent B: pH8.0 10mmol / L Tris-HCl, 2%SDS, 0.1 mmol / L EDTA;
[0038] (3) Reagent C: proteinase K at a concentration of 60 mg / ml;
[0039] (4) Reagent D: It is made by mixing anhydrous phenol and chloroform at a volume ratio of 7:3;
[0040] (5) Reagent E: pH 5.3, sodium acetate with a concentration of 3mol / L;
[0041] (6) Reagent F: Glycogen with a concentration of 20ug / mL.
[0042] The extraction method is:
[0043] (1) Take 1-5 paraffin sections and place them in a 2 mL Eppendorf tube, add 1.5 mL of reagent A and mix well, incubate in a 37 oC water bath for 20 min, centrifuge at 10000 rpm for 6-10 min, discard the supernatant; repeat the above steps once;
[0044] (2) Add 1.5mL 100% ethanol, incu...
Embodiment 2
[0051] Example 2 Using the RNA extracted by the method of the present invention to amplify target genes of different fragment sizes
[0052] The total RNA of paraffin tissue obtained by the method of the present invention is in a diffuse state due to different degrees of degradation. In order to compare the effect of this method on the amplification of genes with different fragment sizes, frozen tissues and paraffin-embedded tissues were selected respectively in this example. The -actin gene was used as the object of amplification, and 13 different fragments of the β-actin gene were amplified. The specific operation is as follows:
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
PUM
Login to view more
Abstract
The invention discloses a novel method and a kit for extracting high-quality RNA (Ribose Nucleic Acid) from a paraffin-embedded tissue. The method comprises the following steps of: firstly, removing paraffin from a sample by using a nontoxic and non-volatile paraffin removing agent to improve the sensitivity of the paraffin-embedded tissue to lysate; secondly, cracking a tissue sample with SDS (Sodium Dodecyl Sulfate)-containing lysate and protease K, wherein the lysate provides a pH environment suitable for cell cracking, so that proteins are further denaturalized and water solubility is enhanced; thirdly, fully denaturalize the proteins with phenol-chloroform mixed liquid, separating the RNA and DNA (Deoxyribose Nucleic Acid) from the proteins, and chromatographing the RNA and the DNA from the mixed liquid to a water layer; fourthly, precipitating the RNA under the coaction of sodium acetate and glucogen; and lastly, dissolving the RNA in RNase-free water. By adopting the kit and the extraction method, the aim of obtaining total RNA with high quality, a long segment and high yield from the paraffin-embedded tissue is fulfilled. The obtained RNA can be applied to medical molecular biology researches, clinical examination and the like.
Description
【Technical field】 [0001] The invention relates to a new method and reagent for extracting high-quality total RNA from paraffin-embedded conventional pathological tissues. 【Background technique】 [0002] The occurrence of diseases is often accompanied by changes in gene expression status. Comprehensive analysis of gene expression by extracting RNA from fresh or frozen tumor tissues is playing an increasingly important role in understanding disease occurrence, classification, and prognosis. However, there are limited sources of fresh or frozen tissue specimens. In contrast, formalin-fixed and paraffin-embedded tissues come from a wide range of sources, and the pathology departments of major hospitals have a large number of archives. On the other hand, this part of the specimens often has good follow-up data, which makes it possible to conduct large-scale clinical retrospective studies using archived wax blocks. [0003] However, compared with fresh tissue, RNA extraction in...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more