13 results about "Ribosomal RNA" patented technology

The invention provides a macro virus group sequencing library construction method for removing host ribosome RNA through an enzyme method and a kit, and belongs to the technical field of biology. The method comprises the following steps: hybridizing total RNA (Ribonucleic Acid) of a sample with a DNA (Deoxyribose Nucleic Acid) probe aiming at host ribosome RNA to form a DNA / RNA hybrid chain; rNase H is used for digesting RNA in the hybrid chain, and DNase I is used for digesting the remaining DNA probe in the system; purifying to obtain virus RNA (Ribonucleic Acid); carrying out fragmentation treatment on the virus RNA; carrying out reverse transcription on the fragmented RNA to synthesize a first chain cDNA, then synthesizing a second chain cDNA, and carrying out terminal repair and dA tail addition to obtain double-chain cDNA; connecting the linker with the dT tail with the double-chain cDNA; and carrying out PCR amplification and purification on the connection product to obtain the macro virus group sequencing library. According to the construction method and the kit, the problem that the full-length sequence of the virus cannot be obtained from a low-load sample can be solved.

The application discloses a LAMP primer group, a kit and a detection method for detecting Pinus armandii rust fungus, and belongs to the technical field of genetic engineering. The application takes the specific nucleotide sequence of the large subunit ribosomal RNA of the Pinus armandii rust fungus as a target gene to design and screen a primer group which is specific and high in sensitivity, and can be used for LAMP detection of the Pinus armandii rust fungus. The LAMP primer group provided by the application has the advantages of high universality, high specificity, high sensitivity and good accuracy, and the detection limit can reach 4.5x10 ‑4 ng / μL, which is 100 times higher than the sensitivity of the LAMP detection of different species in the same genus in the prior art, the detection method can rapidly detect the Pinus armandii rust fungus from spore piles and host plant materials, and is suitable for the detection of Pinus armandii seedling materials in forest fields and customs quarantine ports.

The use of different primer sets in reverse transcription incorporates tags allowing for the selective amplification of cDNA transcribed using the different primers. Primers targeting non-desired RNA sequences such as ribosomal RNA can be used to prevent subsequent amplification of cDNA transcribed from those non-coding fragments. Accordingly effective depletion of non-desired sequences after cDNA amplification can be achieved. Systems and methods of the invention have applications in whole-transcriptome analysis. Non-coding sequence targeting primers can include nucleotide analogs that, when enzymatically processed, prevent subsequent amplification. Library preparation can include single primer isothermal amplification (SPIA) techniques wherein an RNA sequence required for SPIA is included in random primers but is absent from primers targeting non-coding RNA.

This invention discloses a compound seed treatment agent containing metalaxyl-M and tebuconazole, its preparation method, and its uses, belonging to the technical field of seed treatment agents. The suspension of this invention uses metalaxyl-M, tebuconazole, and sodium selenite as the main components, supplemented with emulsifying and dispersing agents, wetting and dispersing agents, antifreeze agents, suspending agents, film-forming agents, and warning colors, and is prepared by adding deionized water to a sufficient volume. Metalaxyl-M targets and inhibits ribosomal RNA polymerase I of oomycete pathogens, while tebuconazole targets and inhibits sterol 14α-demethylase. The two have complementary fungicidal spectra, synergistically controlling maize stalk rot and maize head smut. Sodium selenite, by inhibiting the oxidative stress defense system of pathogenic fungi involving glutathione and thioredoxin, promotes the effects of the two main components and simultaneously remediates selenium-deficient soils. This invention is suitable for the green and efficient control of maize stalk rot and head smut, and also has the function of ecological restoration of selenium-deficient soils.

The application belongs to the technical field of fluorescent probes, and particularly relates to a preparation method and application of an RNA fluorescent probe based on biological orthogonal reaction, and the structural formula of the fluorescent probe is: the application provides a preparation method for generating an RNA activated near-infrared fluorescent probe based on biological orthogonal reaction. First, (E)-2-chloro-3-(hydroxymethyl)cyclohex-1-en-1-methanal and 6-bromo-1-ethyl-2-methylquinolin-1-ium salt are used as raw materials to synthesize a precursor compound beta-Br, and then the beta-Br is rapidly in-situ generated into a fluorescent probe under physiological conditions. After the fluorescent probe generated based on biological orthogonal reaction is reacted with RNA, the fluorescence intensity at the near-infrared region 670-900 nm is enhanced with the increase of the RNA concentration, and the fluorescence enhancement multiple reaches 400. Secondly, the fluorescent probe generated based on biological orthogonal reaction shows high selectivity to RNA, and is not affected by other various ions, active oxygen, RNA bases, biological thiols and amino acids. Finally, the fluorescent probe can be washed-free and fast imaging in living cells, and can be in-situ fast generated into a fluorescent probe in cells, and can be used for monitoring intracellular ribosomal RNA, distinguishing between living cells and dead cells, and real-time monitoring of intracellular oxidative stress.

This disclosure provides methods and compositions to increase the efficiency of targeted coding RNA pseudouridylation by H / ACA snoRNA, by localizing the H / ACA snoRNA more to the nucleoplasm where pre-mRNAs are transcribed and processed into mature mRNAs versus the nucleolus, where H / ACA snoRNAs natively modify ribosomal RNAs.

The application belongs to the technical field of microorganism detection, and particularly relates to detection primers and probes for Euryphallus sinensis and application thereof. The application establishes a real-time fluorescent quantitative PCR method for the pathogen based on TaqMan-MGB probes according to the small subunit ribosomal RNA (SSUrRNA) gene of Euryphallus sinensis, and designs detection primers based on isothermal amplification technology. With the constructed recombinant plasmid standard as a template, the detection sensitivity of the quantitative PCR detection method for the target gene is 9.58 copies / reaction. The detection primers for isothermal amplification can be detected under simple constant temperature conditions, and the color change of the detection product after adding nucleic acid dye can realize on-site rapid diagnosis of the pathogen.

The invention provides a dual-nucleic acid library co-construction method for synchronous detection of plasma ecDNA and related RNA markers. The method comprises the following steps: firstly, extracting total free nucleic acid from the same plasma sample; then carrying out linear DNA digestion treatment; then removing ribosome RNA (Ribose Nucleic Acid); then carrying out reverse transcription reaction by using a chimeric primer with 6-9 random basic groups at the 3'end and a template conversion oligonucleotide sequence at the 5 'end to obtain single-stranded cDNA with universal joints at the two ends; next, the single-stranded cDNA is subjected to cyclization, and annular cDNA is formed; then carrying out rolling circle amplification on the ecDNA and the annular cDNA; and carrying out fragmentation, terminal repair, linker connection and PCR enrichment on the amplification product to obtain the dual-nucleic acid library for high-throughput sequencing. By adopting the method, the collaborative analysis of the DNA level and the RNA level of the trace sample is realized.

Provided herein are methods, compositions, and systems for processing nucleic acids from individual cells or cell populations. Cells are co-partitioned and processed, allowing for analysis of a variety of types of RNA from cells while minimizing ribosomal RNA species. The present invention enables high-throughput measurement of full length RNA from single cells with a streamlined workflow.

A method for identifying exogenous viruses based on metagenomic sequencing, comprising the following steps: S1, a data quality control step, filtering the quality of the metagenomic sequencing raw data of the sample to obtain clean sequencing data; S2, a host removal step, aligning the clean sequencing data with a host reference genome and a ribosomal RNA database, removing the host sequences on the alignment, and obtaining enriched microbial sequences. The method integrates kraken2 species identification, RVDB / nt_core database alignment, virsorter2 / checkV verification and other multiple links to form a multi-dimensional identification system, improve the accuracy and reliability of virus identification, introduce a multi-process parallel processing mechanism to support the synchronous analysis of batch samples, greatly improve the analysis efficiency, adapt to the high-throughput detection needs of large-scale cell banks, biological products intermediates and finished products, and solve the problem of low throughput of traditional methods and existing mNGS methods.

In an example of a method, a primer functionalized, ribosomal RNA (rRNA) depleted sample is generated. The primer functionalized, rRNA depleted sample is combined with a composition to form a mixture, where the composition includes a reverse transcriptase, an RNA nickase, a DNA polymerase with strand displacement activity or 5'-3' exonuclease activity, and a plurality of deoxynucleotide trisphosphates. The mixture is introduced into a flow cell including a substrate, first and second primers immobilized on the substrate, and first and second transposome complexes immobilized on the substrate. A temperature protocol is initiated to form double-stranded complementary DNA (cDNA) from the primer functionalized, rRNA depleted sample and to form partially adapted cDNA fragments from the double-stranded cDNA.

The application discloses OsRNE A gene and application of the gene and a coded protein in rice leaf color regulation and chloroplast development. OsRNE The nucleotide sequence of the gene is shown as SEQ ID NO. 1, and the amino acid sequence of the coded protein is shown as SEQ ID NO. 2. OsRNE The application utilizes CRISPR / Cas9 gene editing technology to knockout the gene and obtain a loss-of-function mutant. The mutation of the gene leads to impaired chloroplast development of rice, affects chlorophyll synthesis, albino of seedlings and death within three-leaf stage, and thus can be used as a marker trait to assist rice molecular breeding at the seedling stage. OsRNE The gene is mainly expressed in leaves, the coded protein is located in chloroplasts, and the chloroplast development and normal growth of seedlings are affected by affecting the metabolism of ribosomal RNA in chloroplasts. Therefore, the application can be applied to molecular genetic breeding of rice leaf color traits, and has important significance for further understanding of a chloroplast development regulation mechanism of rice and yield increase of rice.

This invention discloses a method and apparatus for mining viral micro-interference RNA using multiple algorithms, relating to the field of biological detection. The method includes: removing adapters from an input sequence using multiple adapter removal algorithms, merging and deduplicating the resulting sequences to obtain an adapter-removed sequence; mapping the adapter-removed sequence to a ribosomal RNA database using multiple mapping algorithms in a cascaded manner, outputting a first incompletely mapped sequence; mapping the first incompletely mapped sequence to a host genome using the same mapping method, outputting a second incompletely mapped sequence; mapping the second incompletely mapped sequence to a viral genome using multiple mapping algorithms. If the second incompletely mapped sequence is mapped to the viral genome by any mapping algorithm with a mismatch number ≤ 1, it is determined to originate from a viral sequence. This invention significantly improves the efficiency and accuracy of mining viral micro-interference RNA from massive transcriptome data.