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144 results about "Single-Stranded RNA" patented technology

An RNA that consists of a single strand of ribonucleotides. Because it lacks a complementary strand this form of RNA can adopt unique tertiary structures.

Methods and reagents to detect and characterize norwalk and related viruses

Double-stranded cDNA was synthesized from nucleic acid extracted from Norwalk virus purified from stool specimens of volunteers. One clone was isolated from a cDNA library constructed in a pUC-13 vector after amplification of the cDNA. The specificity of this cDNA (pUCNV-953) was shown by hybridization assays. The cDNA reacted with post (but not pre-) infection stool samples from Norwalk volunteers and with highly purified Norwalk virus, but not with other common enteric viruses such as hepatitis A virus and rotavirus. Finally, the probe detected virus in the same fractions of CsCl gradients in which viral antigen was detected using a specific Norwalk virus radioimmunoassay, and particles were detected by immune electron microscopy. Single-stranded RNA probes derived from the DNA clone after subcloning into an in vitro transcription vector were also used to show that the Norwalk virus contains a ssRNA genome of about 8 kb in size. The original clone was also used to detect additional cDNAs which represent at least 7 kb of nucleic acid of the Norwalk genome. The availability of a Norwalk-specific cDNA and the first partial genome sequence information allow rapid cloning of the entire genome and of establishment of sensitive diagnostic assays. Such assays can be based on detection of Norwalk virus nucleic acid or Norwalk viral antigen using polyclonal or monoclonal antibodies to proteins expressed from the cDNA or to synthetic peptides made based on the knowledge of the genome sequence. Assays using proteins deduced from the Norwlk virus genome and produced in expression systmes can measure antibody responses. Vaccines made by recombinant DNA technology are now feasible.
Owner:BAYLOR COLLEGE OF MEDICINE

Methods for cleaving DNA with nucleotide integrases

The present invention provides new methods, employing a nucleotide integrase, for cleaving single-stranded RNA substrates, single-stranded DNA substrates, and double- stranded DNA substrates at specific sites and for inserting a nucleic acid molecule into the cleaved substrate. One method uses a nucleotide integrase to cleave one strand of a double-stranded DNA substrate. The method comprises the steps of: providing a nucleotide integrase comprising a group II intron RNA having two hybridizing sequences that are capable of hybridizing with two intron RNA binding sequences on the one strand of the DNA substrate, and a group II-intron encoded protein which binds to a first sequence element of the substrate; and reacting the nucleotide integrase with the double-stranded DNA substrate under conditions that permit the nucleotide integrase to cleave the one strand of the DNA substrate and to insert the group II intron RNA into the cleavage site. The method of cleaving both strands of a double-stranded DNA substrate comprises the steps of: providing a nucleotide integrase comprising a group II intron RNA having two hybridizing sequences that are capable of hybridizing with two intron RNA binding sequences on one strand of the substrate, and a group II-intron encoded protein that is capable of binding to a first sequence element and to a second sequence element in the recognition site of the substrate; and reacting the nucleotide integrase with the double-stranded DNA substrate such that the nucleotide integrase cleaves both strands of the DNA substrate and inserts the group II intron RNA into the cleavage site of the one strand. The method for cleaving a single-stranded nucleic acid substrate comprises the steps of: providing a nucleotide integrase having two hybridizing sequences that are capable of hybridizing with two intron RNA-binding sequences on the single-stranded substrate, and a group II intron encoded protein; and reacting the nucleotide integrase with the single stranded nucleic acid substrate for a time and at a temperature sufficient to allow the nucleotide integrase to cleave the substrate and to attach the group II intron RNA molecule thereto.
Owner:THE OHIO STATE UNIV RES FOUND

Double-labelling fluorescence activation type probe and method for detecting DNA by adopting same

The invention discloses a double-labelling fluorescence activation type probe and a method for detecting DNA by adopting the same. The probe is a single stranded RNA which is complementary with a basic group of to-be-detected DNA and has the length of 5-40 basic groups or is a single stranded DNA which is complementary with a basic group of to-be-detected DNA and has the length of 5-40 basic groups; 1/5-4/5 of basic groups in the DNA sequence are RNA basic groups; The two ends of the probe are labeled with a fluorophore and a quenching group. The probe is simple in structure and fluorescence is completely quenched; when the probe hybridizes with the to-be-detected DNA to form DNA/RNA heteroduplex, RNase H hydrolyzes an RNA chain in a DNA/RNA structure in a specificity manner to enable the fluorophore to be away from the quenching group and enable a fluorescence signal to be recovered; simultaneously, the dissociative probe further hybridizes with the to-be-detected DNA, is hydrolyzed by RNase H and generates the fluorescence signal; circulation is performed in such a way, so that the system fluorescence signal is remarkably amplified to realize rapid and high-sensitive detection of the to-be-detected DNA. The probe can be used for quantitative detection of DNA in an amplification product.
Owner:SHAANXI NORMAL UNIV
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