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

Magnetic bead reagent applied to screening super-long fragment nucleic acids and screening method

A screening method and magnetic bead technology, applied in the field of molecular biology, can solve the problems of poor recovery of large fragments of DNA, shorten the time of adsorption, improve the recovery rate of nucleic acid, and achieve good sequencing results

Inactive Publication Date: 2018-12-21
WUHAN FRASERGEN CO LTD
View PDF7 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the technical problems existing in the prior art, the present invention proposes a magnetic bead solution and a screening method suitable for screening ultra-long fragment nucleic acids. For the problem of poor effect, by designing and optimizing the buffer of magnetic beads, the separation and screening of large fragments using magnetic beads can be realized. It is especially suitable for long-fragment single-molecule sequencing technologies such as PacBio and Nanopore, which can greatly improve sequencing data. fragment lengths for better sequencing results

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
  • Magnetic bead reagent applied to screening super-long fragment nucleic acids and screening method
  • Magnetic bead reagent applied to screening super-long fragment nucleic acids and screening method
  • Magnetic bead reagent applied to screening super-long fragment nucleic acids and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The magnetic bead reagent for screening ultra-long fragment nucleic acid in this example includes silica hydroxy magnetic beads with a concentration of 20 ng / μL, Tris-HCl with a concentration of 5 mmol / L, sodium chloride with a concentration of 1 mol / L, and a concentration of 0.1 mol / L sodium laurate, 0.1mol / L guanidine hydrochloride, 0.5mol / L NaN 3 , polyethylene glycol with a volume percentage concentration of 1% and pH=8 with a concentration of 0.1 mmol / L EDTA. The molecular weight of polyethylene glycol is 8000 Daltons; the silica hydroxyl magnetic beads are derived from Beckman's XP magnetic beads, and the magnetic beads are washed with sterile water to completely remove the residual Buffer of the magnetic beads to obtain the silica of this embodiment. Hydroxy Magnetic Beads.

Embodiment 2

[0040] The magnetic bead reagent for screening ultra-long fragment nucleic acid in this example includes silica hydroxyl magnetic beads with a concentration of 350 ng / μL, Tris-HCl with a concentration of 10 mmol / L, sodium chloride with a concentration of 1.6 mol / L, and a concentration of 10 mol. / L sodium laurate, 5.6 mol / L guanidine hydrochloride, 25 mol / L NaN 3 , the volume percentage concentration is 11% polyethylene glycol, pH=8 concentration is 1mmol / L EDTA, the final concentration is 5mmol / L magnesium chloride and the final concentration is 0.5mmol / L cetyl tritetraammonium bromide . The molecular weight of polyethylene glycol is 8000 Daltons; the silica hydroxyl magnetic beads are derived from Beckman's XP magnetic beads, and the magnetic beads are washed with sterile water to completely remove the residual Buffer of the magnetic beads to obtain the silica of this embodiment. Hydroxy Magnetic Beads.

Embodiment 3

[0042] The magnetic bead reagent for screening ultra-long fragment nucleic acid in this example includes silica hydroxyl magnetic beads with a concentration of 800 ng / μL, Tris-HCl with a concentration of 50 mmol / L, sodium chloride with a concentration of 60 mol / L, and a concentration of 20 mol / L. L sodium laurate, guanidine hydrochloride with a concentration of 10 mol / L, and NaN with a concentration of 100 mol / L 3 , the volume percent concentration is 25% polyethylene glycol, pH=8 concentration is 10mmol / L EDTA, the final concentration is 9.5mmol / L magnesium chloride and the final concentration is 3.5mmol / L hexadecyl tritetrakis bromide Ammonium. The molecular weight of polyethylene glycol is 8000 Daltons; the silica hydroxyl magnetic beads are derived from the magnetic beads of Novozyme, and the magnetic beads are washed with sterile water to completely remove the residual Buffer of the magnetic beads to obtain the oxidation of the present embodiment. Silanol magnetic beads....

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

PropertyMeasurementUnit
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to the technical field of molecular biology and particularly relates to a magnetic bead reagent applied to screening super-long fragment nucleic acids and a screening method. Thereagent comprises silicon oxide hydroxyl magnetic beads with the concentration of 20-800ng / uL, Tris-HCl with the concentration of 5-50mmol / L, sodium chloride with the concentration of 1-60mol / L, sodium laurate with the concentration of 0.1-20mol / L, guanidine hydrochloride with the concentration of 0.1-10mol / L, NaN3 with the concentration of 0.1-100mol / L, polyethylene glycol with the volume percent concentration of 1-25% and EDTA (Ethylene Diamine Tetraacetic Acid) with the pH value of 8 and the concentration of 0.1-10mmol / L. The reagent aims at the problem in the prior art that the large-fragment DNA recovery effect is poor in the high-throughput sequencing library construction process, large fragments are separated and screened by using magnetic beads by designing and optimizing the buffer solution of the magnetic beads, the magnetic bead reagent is particularly applicable to the long-fragment single-molecule sequencing of PacBio, Nanopore and the like, the fragment length of the sequencing data can be greatly improved, and better sequencing results can be obtained.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, and more particularly, to a magnetic bead reagent and a screening method suitable for screening ultra-long fragment nucleic acid. Background technique [0002] High-throughput sequencing technology, namely next generation sequencing technology, can sequence and quantify hundreds of thousands to millions of DNA molecules at a time, providing a powerful tool for basic biomedical research and clinical testing. And promote the scientific research and clinical translation of genomics, laying the cornerstone of its position in precision medicine. Before a sample to be tested is sequenced by a high-throughput DNA sequencing machine, a lot of work needs to be done, and the key step is library construction. Library construction includes nucleic acid fragmentation, repair reaction, A addition reaction, ligation reaction, fragment screening and purification, PCR amplification, PCR product...

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): C12N15/10
CPCC12N15/1013
Inventor 方涛
Owner WUHAN FRASERGEN 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

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