摘要:
Disclosed are a method, device kit, and automated system for simple, reproducible, and high-throughput quantification of mRNA from whole blood. More particularly, the method, device, kit and automated system involve combinations of leukocyte filters attached to oligo(dT)-immobilized multi-well plates.
摘要:
The present invention is directed to methods of generating a signal indicative of the presence of a target nucleic acid in a sample by forming a cleavage structure comprising duplex and single-stranded nucleic acid by incubating a sample comprising a target nucleic acid with a thermostable nucleic acid polymerase substantially lacking 5′ to 3′ exonuclease activity and cleaving said cleavage structure with a thermostable Fen nuclease lacking 5′ to 3′ synthetic activity to generate a signal.
摘要:
The invention relates to methods and systems (e.g., computer systems and computer program products) for determining the biological function of uncharacterized cellular constituents, particularly genes and gene products, by using “response profiles,” i.e., measurements of pluralities of cellular constituents in cells having a modified gene or gene product, as phenotypic markers for the gene or gene product. Methods are provided for clustering such response profiles so that similar or correlated response profiles are organized into the same cluster. The invention also provides databases or “compendiums” of response profiles to which the response profile of an uncharacterized gene or gene product can be compared. In one embodiment, steps of the methods comprise comparing the measured response profiles to response profiles stored in the databases or compendiums, and determining the biological function of the response profiles in the databases that are most similar to the measured response profiles.
摘要:
Methods and kits performing reverse transcription and RT-PCR reactions having high fidelity, processivity and DNA polymerase activity are described. The methods involve performance of reverse transcription at an increased temperature with a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof. The kits of the present invention include a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof, a DNA polymerase capable of amplifying cDNA under conditions suitable for polymerase chain reaction, and the reagents necessary to carry out both processes.
摘要:
A method for amplification of a nucleotide sequence characterized by performing PCR amplification using mRNA isolated from a biological sample as a template and using a first primer comprising a poly(T) sequence and an additional nucleotide sequence X thereto and a second primer comprising a poly(T) sequence and an additional nucleotide sequence Y thereto, provided that the nucleotide sequences X and Y in the first and second primers, respectively, have different sequences from each other.
摘要:
One-step RT-PCR methods, compositions and kits for the detection and quantification of small RNAs in a sample are disclosed. The one-step RT-PCR approach involves polyadenylation of a small RNA followed by reverse transcription with a first primer containing a poly(T) sequence and at least two 3′ nucleotides complementary to the 3′ terminal end nucleotides of the small RNA, to produce a cDNA. This may be followed by PCR amplification using the same first primer as the revere primer and a second, forward primer in which a portion of its sequence is complementary to the 3′ terminal end of the cDNA. This may be then followed by detection and/or quantification of the amplified product.
摘要:
The present invention relates to methods for evaluating the cell damaging potential of an agent by determining the ability of the agent to increase messenger RNA release in cells.
摘要:
The invention relates to compositions for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions include an RNA polymerase, a FEN nuclease, and a probe.
摘要:
The present invention relates to compositions and methods for the detection and characterization of small nucleic acid molecules (e.g., RNA (e.g., small RNAs such as micro RNAs (miRNAs) and small interfering RNAs (siRNAs)) and other short nucleic acid molecules). More particularly, the present invention relates to methods for the detection and quantification of RNA expression. The present invention further provides for the detection of miRNA and siRNA variants.