Unlock instant, AI-driven research and patent intelligence for your innovation.

Kit for constructing next-generation sequencing library and construction method thereof

A technology of next-generation sequencing library and construction method, which is applied in the field of kits and construction of next-generation sequencing library, which can solve the problems of storage, inconvenient use, and various component systems, so as to improve the uniformity and accuracy of sequencing , the effect of easy operation

Pending Publication Date: 2022-06-28
3D BIOMEDICINE SCI & TECH CO LTD
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a kit for constructing a next-generation sequencing library and its construction method, which will solve the problems of various component systems, inconvenient storage and use in the existing kits for the next-generation sequencing library, and can improve the uniformity of the library

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
  • Kit for constructing next-generation sequencing library and construction method thereof
  • Kit for constructing next-generation sequencing library and construction method thereof
  • Kit for constructing next-generation sequencing library and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Preparation of the kit

[0047] The present invention is based on the xGen Prism DNA library prep Kit library building kit (namely IDTPrism library building kit) of IDT Company, which mainly includes: End Repair Buffer, End Repair Enzyme, The first adapter buffer (Ligation 1buffer), the first adapter (Ligation1adapter), the first adapter ligase (Ligation 1Enzyme), the second adapter ligase A (Ligation2EnzymeA), the second adapter ligase B (Ligation 2Enzyme B), the first adapter ligase (Ligation 1Enzyme) Two adapter buffer (Ligation2buffer), second adapter (Ligation 2Adapter), UDI primer (xGenUDI PrimerPairs). In addition, IDTE (pH 8.0, 1×TE solution, IDT company), KAPA HiFi HotStart ReadyMix (Roche company), Ampure XP magnetic beads (Beckman company) were purchased.

[0048] The various components in the kit are premixed, and glycerol and / or Tween are added to obtain an end repair mixed system, a first joint mixed system A, a first joint mixed system B, a second joint mi...

Embodiment 2

[0055] The construction of the second-generation sequencing library, the specific steps are as follows (using each mixed system in Example 1):

[0056] Step 1: After the fragmented DNA fragments are purified by magnetic beads, the fragmented DNA is mixed with the end repair mixing system and placed on a PCR machine for reaction;

[0057] In the specific treatment, the samples used in step 1 were physically interrupted by Covaris, and the DNA after the interruption was a small fragment of DNA of about 200 bp. Take 100-200ng of DNA samples for fragmentation, and then purify the fragmented DNA with 1.8X Ampure XP magnetic beads, and quantify with Qubit after purification. Take 20ng of purified fragmented DNA for library construction. The initial amount of library construction is 20ng. Calculate the amount of DNA sampled according to the corresponding DNA concentration. When performing end repair, add 10uL of end repair mixed system to the PCR reaction tube. And add 50uL of fragm...

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 provides a kit for construction of a next-generation sequencing library and a library construction method, the kit comprises a terminal repair mixing system, a first linker mixing system A, a first linker mixing system B, a second linker mixing system and a PCR primer mixing system.The invention further comprises the construction method of the next-generation sequencing library. All component reagents in the commercial library building kit are reasonably combined and premixed, so that the quantity of single components in the kit is reduced, and the operation time is shortened; small-volume components are premixed into large-volume mixed components, so that the accuracy of the reaction addition amount is ensured; by adding the protective agent or the stabilizer, the stability of the mixed components is ensured, and the mixed components can be stored at room temperature for a long time. The kit provided by the invention simplifies library construction operation, is easy to store, and improves the homogeneity of the library.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for constructing a second-generation sequencing library and a construction method thereof. Background technique [0002] High-throughput sequencing technology (High-throughput Sequencing), also known as next-generation sequencing technology (Next-Generation Sequencing), second-generation sequencing technology, is a parallel sequence determination of hundreds of thousands to millions of DNA molecules, which makes it possible The detailed and comprehensive analysis of the genome and transcriptome of a species is also called Deep Sequencing. [0003] Before high-throughput sequencing, the sequencing library must be constructed first. At present, common library construction kit manufacturers include IDT, Roche, NEB, etc. IDT's xGen Prism DNA library prep Kit can effectively reduce the generation of adapter dimers due to its unique adapter connection method, and can detect as low 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
IPC IPC(8): C12Q1/6806C12N15/10C40B50/06
CPCC12Q1/6806C12N15/1093C40B50/06C12Q2525/191C12Q2531/113C12Q2535/122C12Q2521/501
Inventor 张华秦资李芳敏祝君宗玉琦
Owner 3D BIOMEDICINE SCI & TECH CO LTD