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Universal detection kit

A detection kit and kit technology, applied in the field of biomedicine, can solve the problems of increased chances of non-specific amplification, complicated separation procedures, etc.

Inactive Publication Date: 2021-02-23
新开源汇诚(武汉)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high hybridization sensitivity and specificity, and is more convenient to operate, but the ratio of primers needs to be optimized, and the chance of non-specific amplification increases. In addition, the DNA single-strand separation process needs to rely on electrophoresis, and the separation procedure is complicated. All the separation methods mentioned above have certain limitations.

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Embodiment Construction

[0010] Below in conjunction with embodiment the present invention is described in further detail and completely.

[0011] 1 Universal detection kit to prepare single-stranded DNA for pyrosequencing and DNA chip hybridization.

[0012] 1.1 Composition of Universal Detection Kit

[0013] Binding solution: 10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 1 M NaCl, 0.01%-0.1% Tween-20 and 5% agarose microbeads (streptavidin-labeled, 34 μM in diameter).

[0014] Denaturing solution: 1M NaOH solution.

[0015] Annealing solution: 10mM Tris-HCl (pH7.5), 1mM EDTA (pH7.5).

[0016] Washing solution: pH7.4PBS

[0017] 1.2 PCR

[0018] Ten cases of human gDNA were amplified by PCR with primers MTHFR-F: 5'-GCTGACCTGAAGCACTTGAAG; MTHFR-R: 5'-biotin-CCCATGTCGGTGCATGCCTTCAC. 50μl PCR system is as follows

[0019] components volume 10*buffer 5μL dNTPs (2.5mM each) 4μL MTHFR-F (10μM) 1μL MTHFR-R (10μM) 1μL rTaq (5U / μL) 0.2 μL Human Genomic DNA (10ng / ...

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Abstract

The invention discloses a universal detection kit. The kit is used for single-stranded DNA preparation steps of technical platforms such as pyrosequencing and nucleic acid chip hybridization. The kitcomprises a binding solution, streptavidin-labeled agarose microspheres, a denaturing solution, an annealing solution and a washing solution. The kit provided by the invention is simple and convenientto operate, safe, non-toxic, few in components and low in use cost, and the prepared single-stranded DNA can be applied to detection of the technical platforms such as pyrosequencing and nucleic acidchip hybridization and has very wide application prospects.

Description

technical field [0001] The invention relates to a universal detection kit, which belongs to the field of biomedicine. Background technique [0002] DNA single-strand separation technology is one of the most common separation technologies in the field of biomedicine. It is suitable for various scales of DNA sequencing and probe equipment for different nucleic acid samples, and is widely used in biology, environmental monitoring, and medical diagnosis and testing. . Commonly used DNA single-strand isolation methods in the field of biomedicine include thermal denaturation, enzymatic, magnetic bead, and asymmetric PCR, and each method has its own advantages and disadvantages: 1. Thermal denaturation. By heating the PCR product to make it single-stranded, although the principle of the method is feasible and the operation is simple, this method is gradually not used as the purification of single-stranded DNA due to its low separation rate and purity, and is mostly used for the se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6876
CPCC12Q1/6876
Inventor 张辉
Owner 新开源汇诚(武汉)医疗科技有限公司