37 results about "Northern blot" patented technology

Disclosed are methods and apparatus for separation of biomolecules via two-dimensional gel electrophoresis, methods and apparatus for immunoblotting separated biomolecules, and methods for the use of biomolecules processed via the methods and apparatus of the present invention, including use in a clinical setting. The methods and apparatus for separation of biomolecules via two-dimensional gel comprises vertical agarose gel electrophoresis in the first dimension, and the electrophoresis of a novel non-denaturing 3-35% concave gradient polyacrylamide gel in the second dimension. This novel gel can be cast in a modified gel caster that can facilitate the pouring of multiple gels simultaneously. The methods and apparatus for immunblotting are useful with any type of immunoblotting, including Western blot, Northern blot, and Southern blot analyses. These methods and apparatus provide safe, efficient and cost-effective immunoblots, while facilitating the reduction of exposure to toxic or radioactive materials, as well as the disposal of those materials.

The invention discloses a method for detecting miRNAs by a DIG labeling EDC (carbodiimide) cross-linking bridging method. Once a DIG labeling system is utilized, the sensitivity of the DIG labeling system is improved by at least 50 times in contrast with that of a traditional DIG labeling Northern blots detection system; and the sensitivity of the DIG labeling system is equivalent with that of a radioisotope labeling Northern blots detection system; the method cannot cause any pollution on environment, and is very safe and free of a hybridization step; all operations can be completed in 9-10 hours; and therefore, the method is convenient, quick, and low in detection cost. As a result, the problems of low safety, complex procedure, high price, low sensitivity and the like of a current detection method are solved.

The method for detecting gastric cancer comprises detecting canceration of gastric tissue cell by detecting the inactivation of the human VLDLR (VLDL receptor) gene in the gastric tissue cell by (homozygote) deletion, methylation of CpG island, etc. A DNA chip method, a Southern blot method, a Northern blot method, a real time RT-PCR method, an FISH method, a CGH method, etc., may be cited as the detecting means.

This invention describes methods for the generation of nucleic acid probes that improve the sensitivity of hybridization assays. The sensitivity increase results from structural modifications of nucleic acids that promote network formation during hybridization with the result that a single target molecule becomes attached to a complex of many probe molecules. The structural modification involves fragmentation of the probe nucleic acid followed by joining the fragments together such that their order and orientation and number is altered from the original probe molecule. The result is the generation of permuted probe libraries. Various fragmentation and joining methods are described. Labeling can be done by standard methods before during or after formation of permuted probe libraries. Individual members of permuted probe libraries can be isolated and amplified and perpetuated. Fixed mixtures of such isolated library members can be used as permuted probe libraries of limited variability to control network complexity. Libraries can be prepared with additional sequences not present in the target and the fraction of the library made up by such sequences can be controlled. Such extra-target additions can be used as targets for detection by secondary probes. Since their number can be far greater than equimolar with the probe sequences represented in the target they can be detected with higher sensitivity in the networks. Probes for different targets can incorporate different non-target sequences in hyper-molar quantities permitting sensitive detection of multiple hybridization targets in the same sample. Probes made according to this invention can be used in many kinds of hybridization assay including Southern. blots, Northern blots, Dot blots, Nucleic acid Array hybridization, ‘in situ’ hybridization with fluorescent or other labels (FISH) and various kinds of sandwich hybridization assays.

The invention discloses a method for quickly extracting low molecular weight RNA of plant. After cells are disrupted, genomic DNA and high molecular weight RNA form a DNA-protein complex and a RNA-protein complex with protein respectively; and the DNA-protein complex and the RNA-protein complex are insoluble in acid and low-salt environment, and can be removed and extracted through acid and phenol. The low molecular weight RNA stays in supernatant solution, and can be recovered through the precipitation of ethanol or isopropanol. The method is quick and efficient, and can recover the low molecular weight RNA only by one-step precipitation after phenol extraction. The low molecular weight RNA extracted by the method has high purity, and can be used for the cloning of the low molecular weight RNA and the detection of Northern blot hybridization. The method has simple and quick experimental process, and is suitable for the extraction of the low molecular weight RNA of various plant tissues.

Provided are DNA microarray techniques that allow hybridization without RNA amplification, without using cDNA, and without labeling the nucleic acid prior to hybridization. Referred to as the Double-stranded Exonuclease Protection (DEP) assay, the technique permits the sample RNA to be used directly for hybridization, without manipulation in any way. Further provided is a microarray technique for high-throughput miRNA gene expression analyses, termed the RNA-primed, Array-based, Klenow Enzyme (RAKE) assay. The RAKE assay is a sensitive and specific technique for assessing single-stranded DNA and RNA targets, and offers specific advantages over Northern blots.

The invention discloses a kit and method for detecting microRNA (ribonucleic acid)-155. The kit comprises asymmetrical hairpin probes shown in SEQ ID NO.1 and SEQ ID NO.2. Compared with traditional RT-PCR (reverse transcription-polymerase chain reaction), northern blot and other technologies, the kit has the prominent advantages that (1) the kit carries out liquid-phase hybridization reaction, is closer to the natural state of molecular reaction, solves the problem of high steric hindrance of solid-phase hybridization, and improves the hybridization reaction efficiency; (2) the kit has high detection efficiency and high detection speed, and the traditional nucleic acid detection technology usually needs a few hours and even a dozen hours while the technology carries out molecular recognition and cascade signal amplification simultaneously, can complete nucleic acid detection within ten minutes and does not need to carry out RT-PCR on samples in advance; and (3) the reaction system is simple, dispenses with enzyme reaction and is low in cost.