摘要:
The present invention provides methods for the sequencing of all RNA species within an RNA sample, such as the RNA content obtained from a cell, a tissue, a living organism, or from an artificial source. RNA molecules within the samples are labeled in a RNA-specific manner prior to immobilization on a solid support. One label is used to mark the location of the RNA molecule on the solid support, whereas the second label is used to mark selectively the S′ end of full-length mRNA molecules. RNA molecules are sequenced while being bound to the solid support in one or more sequencing reactions, and sequences of individual RNA molecules can be forwarded to computational analysis for assembling sequence information from individual sequencing reads obtained from the same location on the solid support. Not only unsupervised expression profiling on a genome-wide scale, but also the direct analysis of RNA-RNA interactions become possible as revealed by the analysis of the sequencing information obtained along with genomic information.
摘要:
A method is disclosed for the modification of an end of RNA molecules and the use of such modified RNA molecules in cDNA synthesis for the purpose of cloning, detection, sequencing, and amplification of parts of the RNAs, the entire RNAs, or any cDNAs derived from such modified RNAs. The invention relates further to the amplification and the identification of nucleic acid molecules for the purpose of single molecule detection and/or high-throughput sequencing. In addition, a method is provided for the preparation of pooled samples that contains molecules each of which is marked by an “Identifier Sequence” for its origin. The invention facilitates studies on biological systems and analysis of genes expressed therein.
摘要:
A method is disclosed for obtaining the 5′ends of transcribed regions from a plurality of nucleic acid fragments obtained from biological materials or synthetic pools. DNA fragments encoding the 5′ends are enriched for their individual analysis or for the analysis of concatemers thereof. The sequence information derived from 5′ ends can be used for characterization and cloning of the transcriptome.
摘要:
The invention is a method and a kit for preparing single-stranded DNA from double-stranded DNA and the purification of single-stranded DNA derived from double-stranded DNA. A single-stranded-DNA binding substance is used in combination with a double-strand-specific endonuclease for the removal of undesired double-stranded DNA from a single-stranded DNA preparation and for other related purposes.
摘要:
An object of the invention is to provide a method for increasing enzymatic reactivity to a target substance immobilized on a support; and a method for reducing or suppressing an inhibitory effect of a support on enzymatic reaction. The above object is achieved by a method for increasing enzymatic reactivity to a target substance immobilized on a support by allowing at least one substance selected from the group consisting of saccharides, amino acids, polyhydric alcohols and derivatives thereof to exist; and a method for reducing or suppressing an inhibitory effect of a support immobilized with a target substance on enzymatic reactivity to the target substance by allowing at least one substance selected from the group consisting of saccharides, amino acids, polyhydric alcohols and derivatives thereof to exist.
摘要:
The present invention provides a method for identifying, analyzing and/or cloning nucleic acid isoforms comprising the steps of: preparing at least two nucleic acid isoforms, complementary to each other, hybridizing the at least two complementary nucleic acid isoforms and forming double strand RNA/RNA or DNA/DNA hybrids comprising unpaired regions; recovering the RNA/RNA or DNA/DNA hybrids comprising unpaired regions from not hybridized nucleic acids and from nucleic acids not comprising unpaired regions; and identifying, analyzing and/or cloning the recovered nucleic acid fragment comprising unpaired regions.
摘要:
A novel protein which binds with tumor necrosis factor receptor associated factor 2, TRAF2, as well as a nucleic acid coding for the protein is disclosed. From a mouse cDNA library, a cDNA coding for a novel protein which binds with TRAF2 was discovered by mammalian two-hybrid assay, the cDNA was sequenced, the protein encoded by the cDNA was produced, and the fact that the protein binds with TRAF2 was experimentally confirmed.
摘要:
Means to circulate any nucleic acid molecule and to obtain from such circular nucleic acid molecules fragments that mark both ends of the initial nucleic acid molecule are provided. Means of high value to studies including, but not limited to, expression profiling, splicing, promoter identification, identification of genetic elements, and beyond, which are essential components of commercial applications and services including, but not limited to, drug development, diagnostics, or forensic studies are also provided.