Construction method of rRNA silent RNA library and kit

An oligonucleotide and specific technology, used in the preparation of rRNA silencing RNA libraries, reagents or kits, can solve the problems of rRNA loss in integrity requirements, hybridization sites cannot be effectively removed, etc.

Pending Publication Date: 2021-09-07
VAZYME BIOTECH NANJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are clear species limitations in the use of the kit, and all archaeal samples are not applicable; and limited by the type and quantity of probes used, the kit has high requirements for the integrity of RNA samples—degraded rRNA is often lost Hybridization sites cannot be effectively removed

Method used

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  • Construction method of rRNA silent RNA library and kit
  • Construction method of rRNA silent RNA library and kit
  • Construction method of rRNA silent RNA library and kit

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0062] Use Illumina's Ribo-zero rRNA removal kit for rRNA removal and Stranded Total RNA Sample Preparation kits were used for library construction as a control.

[0063] In this example, 1000 ng of total RNA extracted from conventionally cultured HEK293 cells was used as a starting sample to perform rRNA silencing and construct a transcriptome library for sequencing. The specific process is as follows:

[0064] 1. RNA fragmentation and primer hybridization

[0065] 1000ng of total RNA was treated with RNase-free ddH 2 O diluted to 7 μl.

[0066] Prepare the following mixture in an RNase-free centrifuge tube:

[0067] Table 1

[0068]

[0069]

[0070] a Nanjing Nuoweizan Biotechnology Co., Ltd. (vazyme, #N402), including random primers.

[0071] b See Sequence Listing. The concentration of each primer was 50 μM. All primers were modified with a C3 spacer at the 5' end. All A, T, C bases are modified: C, 5-Methyl dC; T, Super T; A, 2, 6-Amino-dA.

[0072] Per...

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Abstract

The invention provides a method for preparing an rRNA silent RNA library and a reagent or a kit used by the method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for preparing an rRNA silenced RNA library and a reagent or kit used therefor. Background technique [0002] High-throughput sequencing technology, also known as second-generation sequencing technology, can be abbreviated as NGS, which refers to the technology of sequencing hundreds of thousands to several million DNA molecules in parallel at a time, and the length of the sequence is generally short. [0003] A major problem with transcriptome sequencing is the presence of interfering RNA molecules. For example, ribosomal RNA (rRNA) is the most abundant molecule in total RNA, where typically more than 90% of total RNA is rRNA. However, ribosomal RNA provides little information about transcription. In applications such as RNA sequencing (RNA-seq), it is of great interest to maximize the amount of information received from a sequencing run. If library construction involves ab...

Claims

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Application Information

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IPC IPC(8): C40B40/06C40B50/06C12Q1/6806C12N15/11
CPCC40B40/06C40B50/06C12Q1/6806C12Q2521/107C12Q2521/101C12Q2525/191
Inventor 曹林聂俊伟瞿志鹏叶廷跃韩锦雄吴恒景雅
Owner VAZYME BIOTECH NANJING
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