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

dna library construction method with low input

A DNA library and construction method technology, which is applied in DNA preparation, recombinant DNA technology, chemical library, etc., can solve the problems that samples are difficult to obtain and cannot meet the needs of large fragment library construction, and achieve the effect of reducing the experimental process

Active Publication Date: 2021-02-26
CHEERLAND BIOTECH CO LTD
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the application of this technology in the detection of clinical chromosomal structural variation, some types of samples may be difficult to obtain or the amount is relatively small, such as tumor puncture samples, amniotic fluid cells, paraffin sections, etc.
At this time, the sample DNA may not be able to meet the needs of conventional large-fragment library construction

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
  • dna library construction method with low input
  • dna library construction method with low input
  • dna library construction method with low input

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0039] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings. In the following implementation manners, many details are described for better understanding of the present application. However, those skilled in the art can easily recognize that some of the features can be omitted or replaced by other materials and methods in different situations.

[0040]In addition, the characteristics, operations or characteristics described in the specification can be combined in any appropriate manner to form various embodiments. At the same time, the steps or actions in the method description can also be exchanged or adjusted in a manner obvious to those skilled in the art. Therefore, various sequences in the specification and drawings are only for clearly describing a certain embodiment, and do not mean a necessary sequence, unless otherwise stated that a certain sequence must be followed.

[0041] The m...

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

A method for constructing a DNA library with a low initial amount, the method comprising: taking genomic DNA and adding random primers labeled with biotin for annealing and performing an amplification reaction; separating and purifying the whole genome amplification product; Fill in to obtain linear blunt-ended DNA; add ligase to the filled-in linear blunt-end DNA for ligation reaction; purify to obtain double-stranded circular DNA, and then break the double-stranded circular DNA into linear DNA fragments; use streptomycin Magnetic beads capture linear DNA fragments labeled with biotin; carry out end repair and A tail base reaction on the captured linear DNA fragments; connect adapters at both ends of the linear DNA fragments after adding A tail base reactions; The product was amplified by PCR to obtain a DNA library. The method uses biotin-labeled random primers to randomly amplify a small amount of genomic DNA, and at the same time realizes the amplification, truncation and biotin labeling of genomic DNA, thereby realizing the construction of a large-fragment DNA library with low initial input.

Description

technical field [0001] The invention relates to the technical field of library construction, in particular to a low-input DNA library construction method. Background technique [0002] High-Throughput Sequencing (High-Throughput Sequencing), that is, Nextgeneration Sequencing technology (Nextgeneration sequencing), is to achieve large-scale parallel sequencing on high-density biochips, which has the characteristics of high data output and low cost per unit of data volume. But its disadvantage is that the sequencing read length is short, and the general sequencing length is 2X300bp or 2X150bp. When the obtained short-read sequences have no reference genome alignment and assembly, or the genome contains highly complex structural sequences, the sequence alignment and assembly will be very difficult. At this time, the splicing and assembly of short sequences can be assisted by a large-span large fragment library (mate pair library). In addition, the link algorithm is used to a...

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 Patents(China)
IPC IPC(8): C12N15/10C12Q1/6806C40B50/06
Inventor 陈建国陈川杨传春张瑜巨庄丽雯
Owner CHEERLAND BIOTECH CO LTD
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