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Clean-CL purifying kit and application thereof

A technology of kits and reagents, applied in the field of Clean-CL purification kits, can solve the problems of increasing the overall human genome detection cost and high price, and achieve the effects of good usability and stability, cost reduction, and guaranteeing the quality of library construction.

Pending Publication Date: 2017-10-10
北京创新乐土生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The high price of existing imported magnetic bead purification reagents increases the overall cost of human genome detection

Method used

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  • Clean-CL purifying kit and application thereof
  • Clean-CL purifying kit and application thereof
  • Clean-CL purifying kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, Clean-CL purification kit and its usage method

[0048] 1. Clean-CL purification kit

[0049]The Clean-CL purification kit of the present invention comprises a Clean-CL magnetic bead purification reagent, and the Clean-CL magnetic bead purification reagent consists of carboxylate magnetic beads SpeedBeadMagneticCarboxylate Modified Particles, 1M Tris-HCL solution (pH8.0), 0.5M EDTA solution , sodium chloride, polyethylene glycol 11000 and sterile purified water. The amount of each component in the 10ml reagent volume of Clean-CL magnetic bead purification reagent is shown in Table 3. The specific configuration method is as follows:

[0050] Table 3. The amount of each component in the Clean-CL magnetic bead purification reagent of 10ml reagent volume

[0051]

[0052]

[0053] (1) Take 205ul of carboxylate magnetic beads Speed ​​Bead Magnetic CarboxylateModifiedParticles and place at room temperature for 30 minutes, during which time vortex every ...

Embodiment 2

[0083] Embodiment 2, the application of Clean-CL purification kit

[0084] Using 16 venous blood samples as experimental materials, the BIOO library construction kit and the Clean-CL purification kit in Example 1 were used to construct libraries in batches, and the Clean-CL magnetic bead purification reagent in the Clean-CL purification kit of the present invention was detected. stability. At the same time, using Beckman Agencourt AMPure XP-nucleic acid purification kit and AxygenAxyPrepTM Mag PCR Clean-up Protocol kit as controls, construct a parallel library and compare the Clean-CL purification kit of the present invention with Beckman Agencourt AMPure XP-nucleic acid purification kit and Axygen The AxyPrepTM Mag PCR Clean-up Protocol Difference. The reaction conditions, reaction reagents, and the amount of magnetic beads used during the library construction test were all the same, and only the type of magnetic beads used for purification was different. Specific steps are...

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PUM

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Abstract

The invention discloses a Clean-CL purifying kit and application thereof. The Clean-CL purifying kit is a magnetic bead method NDA product purifying kit and contains a magnetic bead purifying reagent, wherein the magnetic bead purifying reagent is a solution prepared by uniformly mixing Speed Bead Magnetic Carboxylate Modified Particles, a Tris-HCL solution, an EDTA solution, sodium chloride, polyethylene glycol and water, can be effectively combined with 100bp-500bp segments and is capable of effectively removing dNTP, primers, a primer dimer, joints, saline ions and other impurities. Experiments prove that the magnetic bead purifying reagent in the Clean-CL purifying kit is good in usability and stability and capable of guaranteeing the construction quality of a library and can be applied to the construction of Sanger and NGS libraries, the purification of a PCR system and a coupled reaction system and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Clean-CL purification kit and application thereof. Background technique [0002] Ever since the double helix structure of DNA was resolved, enormous efforts have been made to understand the complexity and variability of the genome in health and disease. In order to support the smooth progress of the Human Genome Project, great improvements have been made in instruments and reagents. The completion of this program has made people strongly aware that people need more and better technology and data analysis capabilities to answer a series of biological questions that follow. However, the limitation of throughput and the high cost of sequencing have become a hurdle for people to further understand the genome. The high-throughput sequencing platform launched after 2000 has solved this problem well, and the cost of human genome sequencing has directly dropped by 50,000 time...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 王金龙张瑞吕灵双石太平宋金良刘如银
Owner 北京创新乐土生物科技有限公司