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Micro-platelet RNA sequencing detection kit based on BGISEQ-500

A technology of platelets and kits, applied in the biological field, can solve the problems of low detection accuracy, limited number of detection genes, large sampling volume, etc., and achieve high sensitivity and high specificity

Inactive Publication Date: 2019-06-25
SHENZHEN HUADA GENE INST
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Problems solved by technology

The shortcomings of platelet protein detection are: large sampling volume and low detection accuracy; the shortcomings of platelet RNA quantitative PCR detection are: limited number of detection genes and low detection accuracy

Method used

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  • Micro-platelet RNA sequencing detection kit based on BGISEQ-500
  • Micro-platelet RNA sequencing detection kit based on BGISEQ-500
  • Micro-platelet RNA sequencing detection kit based on BGISEQ-500

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Embodiment 1

[0061] Example 1. Microplatelet RNA sequencing method and application based on BGISEQ-500

[0062] The present invention is mainly based on the BGISEQ-500 sequencer to sequence and analyze the platelet RNA of 8 normal samples and 15 cancer samples, and the specific steps are as follows:

[0063] (1) Micro platelet RNA library construction

[0064] 1. Platelet separation

[0065] Collect 2ml of peripheral blood from the sample to be tested in an EDTA anticoagulant tube, mix well and perform the following operations. All centrifugation processes are carried out at 4°C:

[0066](1) Place the blood collection tube in a centrifuge and centrifuge at 120g for 20min. Immediately after centrifugation, divide the supernatant (containing platelets) into multiple 1.5ml centrifuge tubes;

[0067] (2) Place the centrifuge tube containing the supernatant obtained in (1) in a centrifuge and centrifuge at 360g for 20min. After centrifugation, the precipitate in the tube is platelets. Add 30u...

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Abstract

The invention discloses a micro-platelet RNA sequencing detection kit based on BGISEQ-500. The invention also provides a method for constructing a platelet RNA sequencing library based on BGISEQ-500.The method comprises the following steps: 1) extracting platelet RNA from a sample to be detected, and performing reverse transcription to obtain dscDNA; 2) fragmenting the dscDNA by a linker-embeddedTn5 transposase to obtain a fragmented product; 3) carrying out PCR amplification of the fragmented product to obtain an amplification product; and 4) cyclizing the amplification product to obtain the platelet RNA sequencing library. Experiments prove that below 1 ml of whole blood is adopted to construct the micro-platelet mRNA library, and an autonomously established platelet analysis procedureis used to carry out highly-sensitive and highly-specific early screening of tumors.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a microplatelet RNA sequencing kit based on BGISEQ-500. Background technique [0002] Cancer is the leading cause of morbidity and mortality worldwide, with approximately 14 million new cancer cases and 8.2 million cancer-related deaths worldwide in 2012. In 2012, the number of cancer cases in China was 3.065 million, accounting for about 20% of the global cancer cases, ranking first in the world; the number of cancer deaths was 2.205 million, accounting for about 1 / 4 of the global cancer deaths. In the next 20 years, it is estimated that the annual global cancer cases will rise from 14 million in 2012 to 22 million. The treatment of malignant tumors is difficult and the medical expenses are high. Many patients have reached the advanced stage when their cancer is discovered, and have lost the best time for treatment. Therefore, early diagnosis of tumor plays an extremely important...

Claims

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Application Information

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IPC IPC(8): C40B50/06
Inventor 陈小芳谢颖周清易吉李南南罗慧娟于竞朱师达
Owner SHENZHEN HUADA GENE INST
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