Library building method for sRNA sequencing

A sequencing library and sequencing technology, applied in the field of small RNA library construction, can solve problems such as increased mismatch rate and low detection rate

Pending Publication Date: 2021-08-13
成都罗宁生物科技有限公司
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  • Abstract
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Problems solved by technology

This method was later shown to have a lower detection rate for miRNAs compared to adapter ligation, resulting in increased mismatch rates

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  • Library building method for sRNA sequencing
  • Library building method for sRNA sequencing
  • Library building method for sRNA sequencing

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Embodiment Construction

[0027] Embodiments of the present invention are described in detail below, examples of which are shown in the drawings, wherein the same or similar reference numerals designate the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by referring to the figures are exemplary and are intended to explain the present invention and should not be construed as limiting the present invention.

[0028] In describing the present invention, it should be understood that the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", The orientation or positional relationship indicated by "horizontal", "top", "bottom", "inner", "outer", etc. are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than Nothing indicating or implying that a referenced device or element...

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Abstract

The invention discloses a library building method for sRNA sequencing. The library building method comprises the following steps that firstly, an obtained sample is pretreated, and RNA is extracted and prepared; then, a 3'end connector and a 5 'end connector which are subjected to pre-adenylation are pretreated, and the 3' end connector and the 5 'end connector are connected with the prepared RNA and purified; reverse transcription reaction is carried out on the purified connection product, meanwhile, the obtained corresponding cDNA chain is purified, and PCR enrichment reaction is carried out on the cDNA chain to obtain a PCR product containing adapter; and finally, the PCR products are subjected to size sorting and purification according to the requirements of the sRNA library, the sRNA high-throughput sequencing library is obtained, and the mismatch rate is reduced.

Description

technical field [0001] The invention relates to the technical field of small RNA library construction, in particular to a library construction method for sRNA sequencing. Background technique [0002] Small RNAs (sRNAs) are a type of RNA that play an indispensable role in gene regulation at the transcriptional and post-transcriptional levels. There are various techniques applied to the quantification of sRNAs, including hybridization, qPCR, and high-throughput sequencing (NGS). NGS technology is undoubtedly a low-cost method of choice for genome-wide quantification of sRNA. At the same time, NGS is also the only technology that can discover new sRNA sequences, distinguish similar sRNA classes, and confirm post-transcriptional modification sequences. The traditional sRNA library construction workflow includes: 1. Add adapters for single-stranded RNA ligation; 2. Reverse transcription; 3. PCR. Most studies have shown that linkage is the cause of deviations in the entire lib...

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Application Information

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IPC IPC(8): C12Q1/6806C12Q1/6869C40B50/06
CPCC12Q1/6806C12Q1/6869C40B50/06C12Q2535/122
Inventor 丁泽琴孙梓健涂波
Owner 成都罗宁生物科技有限公司
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