Efficient and accurate high-throughput detection sample quality evaluation method

A measurement method and sample technology, applied in the field of high-throughput sequencing, can solve the problems of high cost of QPCR reagents, long time required, and high operation requirements, and achieve the effects of shortening evaluation time, saving time, and low technical requirements

Active Publication Date: 2019-01-18
GUANGZHOU BURNING ROCK DX CO LTD
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) QPCR judgment fragment: complex operation and time-consuming: QPCR method is a more accurate method than electrophoresis, but because of its high operation requirements, complicated operation and long time, 3 to 4 hours, this clinical report cycle brings Very high pressure, so it has not been widely used; economy: the cost of reagents for QPCR is relatively high

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  • Efficient and accurate high-throughput detection sample quality evaluation method
  • Efficient and accurate high-throughput detection sample quality evaluation method
  • Efficient and accurate high-throughput detection sample quality evaluation method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 N / Q ratio evaluation method

[0040] 1. Experimental operation

[0041] 1. Extraction of sample DNA

[0042] DNA was extracted using a commercially available tissue or cell DNA extraction kit.

[0043] 2. Determine the interruption conditions and perform mechanical interruption

[0044] Use the Nanodrop spectrophotometer and the Qubit fluorometer to measure the DNA concentration of the sample respectively (use the Qubit dsDNA HS Assay Kit with product number Q32854 in combination with the Qubit fluorometer for DNA concentration detection), and calculate the concentration ratio according to the following formula:

[0045] N / Q ratio = sample DNA concentration measured using a Nanodrop spectrophotometer / sample DNA concentration measured using a Qubit fluorometer.

[0046] See Table 1 for sample grading labeling and interruption conditions.

[0047] Table 1:

[0048]

[0049]3. Interrupt fragment analysis

[0050] After that, the normal NGS process wil...

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Abstract

The invention discloses an efficient and accurate high-throughput detection sample quality evaluation method. According to the method for evaluating DNS sample degradation condition disclosed by the invention, sample DNA concentration is respectively measured through utilization of a fluorescent dye method and an ultraviolet absorption method, and a specific value of the two measurement methods iscomputed. The accuracy of the method for evaluating the DNS sample degradation condition disclosed by the invention can satisfy follow-up NGS ("Next-generation" sequencing) or even other demands forfragment evolution. Compared with conventional methods, the method has the advantages that time and space are reduced, economical efficiency is high and a human error is low, so the conventional high-throughput sequencing DNA fragment evaluation methods can be replaced by the method provided by the invention. A quality control link and a detection period of high-throughput sequencing can be greatly reduced. The method has great advantage over cost and is deserved to be widely popularized and applied.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, and more specifically, to an efficient and accurate quality assessment method for high-throughput detection samples. Background technique [0002] High-throughput sequencing technology, also known as "Next-generation" sequencing technology (NGS), can simultaneously sequence millions of DNA molecules. volume, scalability, and speed, enabling researchers to study biological systems at unprecedented levels. NGS technology can rapidly sequence entire genomes, amplify for deep sequencing of target regions. High-throughput sequencing technology has been widely used in all aspects of life sciences. However, the NGS testing process is relatively long, starting with sample collection, pathological evaluation, nucleic acid extraction, library building, computer testing, and data analysis. In order to ensure high-quality and accurate results for each clinical test sample, it is necessa...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N21/64G01N21/33
CPCG01N21/33G01N21/6428G01N2021/6439
Inventor段飞蝶韩维汉雨生揣少坤
OwnerGUANGZHOU BURNING ROCK DX CO LTD