Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Novel methylation library construction method and application thereof

A methylation and genome technology, applied in the field of molecular biology, can solve problems such as decreased link efficiency, expensive enzymes, and no linking ability

Inactive Publication Date: 2018-06-15
SHANGHAI MAJORBIO BIO PHARM TECH
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] The method of self-ligation into a circle requires the use of circligase. First, the enzyme will preferentially perform intramolecular circularization, so if it is necessary to access adapters for pre-amplification, the relative linking efficiency will be greatly reduced.
Second, the enzyme is sequence-selective for the ligation of target molecules. It has no ligation ability for molecules with C at the 3' end, and the ligation efficiency for molecules with A\T\C at the 5' end is much lower than that of 5' Molecules that end in a G, so there is a severe bias
Third, the enzyme is expensive

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
  • Novel methylation library construction method and application thereof
  • Novel methylation library construction method and application thereof
  • Novel methylation library construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0225] 1. Test preparation

[0226] (1) Synthesis of cohesive end adapters to be used in the method of methylation library construction:

[0227] The sticky end adapter is a double-stranded linker with a sticky end, one end of the sticky end adapter is a first sticky end, and the first sticky end includes at least a random sequence.

[0228] 1. As an example, use Primer1 and Primer2 to synthesize sticky end adapters,

[0229] Primer1: 5'-AGATCGGAAG-3'-bioten (SEQ ID NO.1);

[0230] Primer2: 5'-CTTCCGATCTNNNNNN-3' (SEQ ID NO.2); wherein, NNNNNN represents a random sequence, usually the length of the random sequence is 2-30nt, preferably 4-20nt, more preferably 5-10nt .

[0231] 2. Add the configuration reaction solution according to Table 1:

[0232] Table 1

[0233] Reagent

Volume (ul)

Primer1 (10μM)

1

Primer2 (10μM)

1

TE buffer

7

5M NaCl

1

total capacity

10

[0234] The TE buffer used in the reaction ca...

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

No PUM Login to View More

Abstract

The invention relates to the technical field of molecular biology and in particular relates to a novel methylation library construction method and an application thereof. The novel methylation libraryconstruction method provided by the invention is suitable for single-strand connection, amplification or library construction of a DNA sample treated by using bisulfite. Due to the adoption of a method for bisulfite treatment and then adhesive end joint connection, sample information loss caused by DNA breakage resulted from drug treatment is greatly reduced.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a novel methylation library construction method and its application. Background technique [0002] Methylation is one of the most common epigenetic modification methods, which regulate gene expression without changing the nucleic acid sequence. DNA methylation usually refers to the addition of a methyl group to the fifth carbon atom of cytosine under the mediation of DNA methyltransferase to make it into 5-methylcytosine (5-mC ). Methylation modification plays an important role in gene imprinting, gene expression regulation, X chromosome inactivation, cancer occurrence and development, etc. [0003] Abnormal DNA methylation can participate in the formation of tumors by affecting the expression of proto-oncogenes and tumor suppressor genes. When tumors occur, tumor suppressor genes usually undergo overall hypermethylation, especially in the CpG island region, thereby ...

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
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/06C12N15/10C12Q1/6869C12Q1/6858
CPCC12N15/1093C12Q1/6858C12Q1/6869C40B50/06C12Q2523/125C12Q2525/191C12Q2521/501
Inventor 陈昌岳李静王芳闫丽郑冠涛路远胡秋萍张祥林
Owner SHANGHAI MAJORBIO BIO PHARM TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com