Sequencing library, preparation method thereof, and terminal sequencing method and device

A technology for sequencing library and terminal sequencing, applied in the field of molecular biology, can solve the problems of limited large-scale promotion, lack of vector versatility, and inability to sequence BAC terminals.

Active Publication Date: 2011-02-09
BGI TECH SOLUTIONS
View PDF3 Cites 37 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the difficulty of assembly, the ends of large fragments need to be sequenced. A method is disclosed in WO2010003316A1, which synthesizes vectors for constructing fosmid clones, and the endonucleases FspBI and Csp6I that recognize 4 bases exist in these vectors The cleavage site of the fosmid was mutated, and these two endonucleases were used to digest the fosmid clone inserted with the exogenous fragment, and the target cleavage fragment was recovered. After circularization, the two terminal sequences of the fosmid clone were obtained, and t...

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
  • Sequencing library, preparation method thereof, and terminal sequencing method and device
  • Sequencing library, preparation method thereof, and terminal sequencing method and device
  • Sequencing library, preparation method thereof, and terminal sequencing method and device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: use the comparison of sequencing method of the present invention and prior art method

[0060] 1) Random interrupt

[0061] Take the polar bear genome DNA (this laboratory uses the salting out method to extract, the specific method can refer to Lahiri D, Schnabel B.1993. DNA isolation by a rapid method from human blood samples: effects of MgC12, EDTA, storage time, and temperature on DNA yield and quality.Biochem Genet.31:321328), ensure that the DNA size is not less than 36Kb, use CopyControl TM HTP Fosmid Library Production Kit (Epicentre, USA), prepared the polar bear fosmid clone library according to the manufacturer's detailed instructions, and extracted plasmid DNA from the fosmid clone mixture by using the alkaline lysis method commonly used in the art.

[0062] Using a standard Hydroshear instrument (GeneMachine, San Carlos, CA., USA), a Custom Shearing Assembly-large (4Kb-40Kb) device (GeneMachine, San Carlos, CA., USA), mix the fosmid clone pla...

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 belongs to the field of molecular biology, and relates to a sequencing library, a preparation method thereof, a terminal sequencing method and a terminal sequencing device. The method for preparing the sequencing library comprises the following steps of: breaking randomly; repairing terminals; separating; cyclizing and amplifying. The terminal sequencing method comprises a step of sequencing by using the sequencing library prepared by the method for preparing the sequencing library, and is particularly suitable for high throughput sequencing of large segments of DNA. The invention also relates to a high throughput terminal sequencing device, which comprises the following units: 1) a random breaking unit, 2) a blunt-end modification unit, 3) a separation unit, 4) a cyclizing unit, 5) a PCR amplification unit and 6) a sequencing unit. The terminal sequencing method overcomes enzyme cutting bias and can obtain longer assembled segments.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a sequencing library and a preparation method thereof, as well as a terminal sequencing method and device. The terminal sequencing method and device are particularly suitable for high-throughput sequencing. Background technique [0002] In genome sequencing, genomic DNA is usually cloned into a vector and then sequenced. For example, commonly used vectors are Fosmid and bacterial artificial chromosome (BAC), both of which have the characteristics of large and stable insert fragments and are important tools for genomics research. It is known that BAC can usually insert a fragment of 100-200kb, and Fosmid can usually insert a fragment of about 40kb. Both of them play an important role in gene map cloning, gene analysis, structural variation and genome assembly. In addition, a variety of other vectors are also used in sequencing. [0003] In the first-generation sequencing technolo...

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
IPC IPC(8): C12Q1/68C12N15/10C40B40/08C40B50/06
CPCC12N15/1093
Inventor 韩长磊徐讯
Owner BGI TECH SOLUTIONS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap