8 results about "Rna degradation" patented technology

A protection method, a protection sequence, a composition and a kit for preventing RNA degradation, and the use thereof, which can improve the protective effect on viral RNA. After the protection sequence RNase AP is added, the three-dimensional structure of RNA and the structural allosteric characteristics of RNA can be protected; and the protection sequence has a good protection effect on RNA integrity, and a high-level detection rate can be obtained by a one-time amplification, thereby ensuring efficient enrichment of viral RNA and providing guarantee for subsequent reverse transcription and PCR amplification.

This disclosure provides several ribonuclease R variants and their preparation methods. Compared to the parent enzyme, the variants described in this disclosure exhibit increased protease activity per unit and show promising prospects for industrial applications. Furthermore, this disclosure also provides the application of ribonuclease R variants in linear RNA degradation.

The invention discloses a single cell whole genome methylation and transcriptome biomics library construction method, and relates to the technical field of sequencing methods.The method comprises the steps that a single cell is added into a sorting tube with a preset lysis buffer for lysis, a nucleophilic coating is arranged on the inner wall of the sorting tube to enable cell nuclei to be adsorbed, and supernate containing mRNA is obtained; carrying out reverse transcription on supernate to obtain cDNA (complementary deoxyribonucleic acid) and constructing a transcriptome library; splitting the cell nucleus to release DNA (Deoxyribonucleic Acid) and carrying out hydrosulfite conversion; the transformed DNA is connected with a molecular tag-containing joint to obtain a methylated intermediate product, after exonuclease treatment, the intermediate products with different molecular tags are combined, and the methylated library is obtained through magnetic bead purification, single chain, connection with a sequencing joint at the other end, amplification and purification. According to the technical scheme, RNA and DNA are separated in the same tube and a library is established in parallel, separation and transfer steps are reduced, RNA degradation and nucleic acid loss are reduced, and methylation library establishment flux and biomics data consistency are improved.

The invention belongs to the technical field of microorganism preservation, and particularly relates to normal-temperature preservation liquid for group B streptococcus (GBS) in a vaginal swab as well as a preparation method and application of the normal-temperature preservation liquid. The invention provides a GBS normal-temperature preservation solution. The formula of the GBS normal-temperature preservation solution comprises glycerol, glucose, sodium citrate, EDTA, FBS, compound amino acid, NaCl and the like. The preservation solution ensures that GBS can be stably preserved for 30 days at normal temperature, the activity of GBS and the stability of RNA are effectively maintained, the problems of RNA degradation and bacterial activity loss in an existing preservation method are solved, and reliable sample support is provided for subsequent bacterial culture and RNA detection. The method is suitable for long-time preservation and subsequent detection of clinical vaginal swab samples, and has a good application prospect.

The application discloses a spherical nucleic acid preparation method based on a cyclic template strategy and application, and comprises the following steps: A) using controllable free radical polymerization (ATRP) and intramolecular click chemistry of copper-catalyzed alkyne and azido, a cyclic polymer is prepared c -P(HEMA) 30 B) using continuous esterification and substitution reaction, the cyclic polymer side chain terminal hydroxyl group is substituted by azido, and a cyclic template c -P(HEMA-N3) 30 C) using copper-free click chemistry reaction of a dibenzo cyclooctyne group (DBCO) and azido, nucleic acid is covalently coupled to the cyclic template, and a cyclic brush macromolecule c -P(HEMA-RNA) 30 The obtained cyclic brush macromolecule is subjected to water dialysis, and spherical nucleic acid is prepared through self-assembly. The spherical nucleic acid prepared by the preparation method has good reproducibility, good in-vitro stability, can better protect the nucleic acid from degradation of RNA degradation enzyme, and can greatly improve the nucleic acid uptake capacity of cells.

The application discloses a kit and a method for extracting nuclei in plant fruit kernels. The kit comprises a lysis solution, a washing solution and a purification solution. The lysis solution comprises 0.04%-0.4% w / v Triton X100, 0.2-0.4 mM spermine, 0.5-1 mM spermidine, 0.4-1.6 U / muL RNase inhibitor, 1-2.56 mM DTT, 10-15 mM NaCl, 1X PBS and 200-350 mM sucrose. The washing solution comprises 2% w / v BSA, 1.25-2.5 mM DTT and 0.2-0.4 U / muL RNase inhibitor. The purification solution comprises methanol and at least two Percoll solutions with different concentrations. The kit with specific components and the optimized extraction method can quickly obtain a high-purity nucleus suspension without extracting protoplasts, effectively remove impurities and secondary metabolites, reduce the risk of RNA degradation, and are simple to operate and low in cost.

This invention discloses a method for constructing a dual-omics library based on single-cell whole-genome methylation and transcriptomics, belonging to the field of sequencing technology. The method includes: lysing single cells in a sorting tube pre-filled with lysis buffer; applying a nucleophilic coating to the inner wall of the sorting tube to allow cell nuclei to adsorb and obtain a supernatant containing mRNA; reverse transcribing the supernatant to obtain cDNA and constructing a transcriptome library; lysing the cell nucleus to release DNA and converting it to bisulfite; ligating the converted DNA with a molecularly tagged adapter to obtain methylation intermediates; treating with exonucleases; merging intermediates with different molecular tags; purifying with magnetic beads, single-stranding, ligating the other end with a sequencing adapter, and amplifying and purifying to obtain a methylated library. This invention achieves simultaneous separation of RNA and DNA in the same tube for library construction, reducing separation and transfer steps, minimizing RNA degradation and nucleic acid loss, and improving the throughput of methylation library construction and the consistency of dual-omics data.