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Method, device and application for preparing candidate sequencing probe set

A probe set and probe technology, applied in biochemical equipment and methods, biochemical cleaning devices, enzymology/microbiology devices, etc., can solve problems such as unable to decode annotations of genetic information, high data bias, and need to be improved , to achieve the effect of accurate and reliable transcript sequence and low data bias

Active Publication Date: 2021-06-08
MGI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current transcriptome sequencing technology has low accuracy of sequencing results and high data bias. It is impossible to decode and annotate more complex genetic information in the future, and it is difficult to analyze transcripts and variable splicing.
[0003] Therefore, the current transcriptome sequencing technology still needs to be improved

Method used

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  • Method, device and application for preparing candidate sequencing probe set
  • Method, device and application for preparing candidate sequencing probe set
  • Method, device and application for preparing candidate sequencing probe set

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] 1. Construction of transcriptome library

[0084] 1. Annealing of Riboprobes to Total RNA

[0085] 1) Take 200ng-5μg total RNA sample (MAQC standard) in an RNase-free 0.2ml PCR tube.

[0086] RNA 200ng-5μg Hybridization probe (10μM) 2μL 5× hybridization buffer 1μL water (nuclease free) Make up to a total volume of 5 μL

[0087] Wherein, the hybridization probe used is the probe used in Example 1 of the patent application with the application number CN 201410505793.2. For the specific sequence, please refer to the description of the patent application, which is hereby incorporated herein in its entirety.

[0088] 2) 95°C, 2min; gradient cooling 0.1°C / sec; 22°C, 5min.

[0089] 3) After the reaction is completed, quickly place it on ice for the next step of reaction.

[0090] 2. RNase H enzyme digestion

[0091] 1) Prepare the reaction mixture according to the following ratio:

[0092]

[0093] 2) React for 30 minutes at 37°C. ...

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Abstract

The invention discloses a method, device and application for preparing a candidate sequencing probe set, wherein the method for preparing a candidate sequencing probe set includes: (1) designing a probe based on a target mRNA sequence of a reference genome, and constructing a candidate probe set; (2) compare the candidate probe set with the target mRNA sequence of the reference genome; (3) screen all the candidate probes in the candidate probe set based on the comparison results; (4) target the target mRNA in the reference genome The same probes are designed for highly homologous genes; (5) Merge specific probe sets and probes for highly homologous genes. This method can effectively obtain a set of candidate sequencing probes for all mRNAs of the reference genome, and then, based on it, can effectively prepare and obtain a transcriptome library-specific sequencing primer set, use the sequencing primer set to perform transcriptome sequencing, and determine the results of the sequencing. The transcript sequence is accurate and reliable, and the data bias is low.

Description

technical field [0001] The present invention relates to the technical field of transcriptome library sequencing analysis, in particular to a method, device and application for preparing candidate sequencing probe sets. Background technique [0002] At present, in the field of transcript assembly library and sequencing, the information analysis of the whole transcriptome can be carried out based on short paired-end paired read length sequences, including the analysis of genetic expression events such as alternative splicing. However, the current transcriptome sequencing technology has low accuracy of sequencing results and high data bias. It is impossible to decode and annotate more complex genetic information in the future, and it is difficult to analyze transcripts and variable splicing. [0003] Therefore, the current transcriptome sequencing technology still needs to be improved. Contents of the invention [0004] The present invention aims to solve at least one of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6806C12Q1/6869C12N15/11C40B40/06C12M1/34C40B50/06
CPCC12Q1/6806C12Q1/6869C40B40/06C40B50/06
Inventor 徐讯蒋慧耿春雨范广益梁恩靖祝珍珍
Owner MGI TECH CO LTD