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320 results about "Full length cdna" patented technology

Method of synthesizing cdna

A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA / cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA / cDNA heteroduplex and the double-stranded DNA primer by joining the 3' and 5' ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.
Owner:KOKURITSU SHINTAI SHIYOUGAISHI +2

Method of preparing normalized and/or subtracted cDNA

A method of preparing normalized and / or subtracted cDNA; a method in which the cDNA that is normalized and / or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and / or subtracted cDNA comprising the steps of: (a) preparing cDNA tester; (b) preparing normalization and / or subtraction RNA driver; (c) conducting normalization and / or subtraction in two steps in any order, or conducting normalization / subtraction as a single step and mixing the normalization / subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester / driver hybrids; and (f) recovering the normalized and / or subtracted cDNA; and a method of efficiently preparing normalized and / or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.
Owner:YOSHIHIDE HAYASHIZAKI +2

Methods and compositions for diagnosis and treatment of cancer

InactiveUS20020086812A1Reducing tumorigenicityReduce dosageOrganic active ingredientsBiocideDiseaseNovel gene
The present invention relates to a novel gene, CaSm, that is highly expressed in cancer tissues and cell lines, especially pancreatic cancer. The full length cDNA of CaSm encodes a protein of 133 amino acids. The present invention further encompasses CaSm peptides, fusion proteins, host cell expression systems, antibodies to CaSm, antisense CaSm molecules, and compounds that modulate CaSm gene expression or CaSm activity. The present invention also encompasses methods for disease diagnosis, drug screening and the treatment of cancer. In particular, the combined use of a CaSm antagonist with a therapeutic agent to treat cancer is encompassed.
Owner:THE MEDICAL UNIV OF SOUTH CAROLINA

Method for extensively screening scallop SNP

InactiveCN101845489AThe principle is safe and reliableReliable Large-Scale Screening RunsMicrobiological testing/measurementMolecular geneticsPhenol
The invention belongs to a method for extensively screening scallop SNP in the technical field of the scallop DNA molecular genetic marker, which comprises the following steps that: solution D and phenol-chloroform are firstly used for extracting a plurality of Chlamys farreri RNA, and a ultraviolet specrophotometer is used for measuring the concentration of the mixture of solution D and the phenol-chloroform which are uniformly mixed; a scallop full-length cDNA sample is re-established and comprises the synthesis of a cDNA first chain and the synthesis and augmentation of a cDNA second chain; then homogenization of full-length cDNA is performed, and ultrasonic breaking is performed; then sequence measuring joints are connected, and terminal repair, joint connection and sample augmentation are included; finally a biological software is used for analyze the data to obtain an SNP marker; then an SNP marker to be screened is selected to design a primer; and DNA of individual scallop is extracted to be equivalently mixed to be used as a template, conventional colony sequence measuring is undertaken after the PCR augmentation, and the position point with the occurring frequency of single base difference being 10 percent or more is defined as an SNP marker. The method has safe and reliable principle, simple and controllable flow procedures, reliable running of large-scale screening, excellent effect and strong practicability.
Owner:OCEAN UNIV OF CHINA

Method for cultivating transgenic plant with improved insect resistance by using RNA interference technology and special DNA fragment thereof

The invention discloses a method for cultivating a transgenic plant with improved insect resistance by using RNA interference technology and a special DNA fragment thereof. The DNA fragment is a gene fragment shown as a formula I, wherein a forward sequence (SEQ) is any one fragment which at least comprises 19bp in full-length cDNA fragments of an ATP synthase E subunit gene; a reverse sequence (SEQ) and the forward sequence (SEQ) are mutually reversely complementary; X is a spacer sequence between the forward sequence (SEQ) and the reverse sequence (SEQ), and the X and the forward sequence (SEQ) or the X and the reverse sequence (SEQ) are both not complementary mutually; and the full-length cDNA of the ATP synthase E subunit gene is shown as a sequence 1 in a sequence table. The invention also aims to provide application of the ATP synthase E subunit gene serving as an RNA interference target gene in cultivating the transgenic plant with improved insect resistance. Experiments prove that aphids inoculated to wild-type tobacco grow and reproduce normally; and aphids inoculated to transgenic tobacco starts to die on the fourth day, dead aphid bodies is blackened after five days and no live aphids can be seen after seven days. The formula (I) is forward sequence (SEQ)-X- reverse sequence (SEQ).
Owner:INST OF MICROBIOLOGY - CHINESE ACAD OF SCI

Preparation method and application of molecular marker of rape male sterile restoring gene

The invention discloses a preparation method and an application of a molecular marker of a rape male sterility restoring gene. The preparation method comprises the following steps of A, designing degenerate primers according to PPR gene conservative amino acid sites, B, extracting a lamina DNA of a rape cytoplasm male sterility restoring line through a cetyltrimethyl ammonium bromide (CTAB) method and magnifying the lamina DNA through a touchdown polymerase chain reaction (PCR) process to obtain a specific restoring gene candidate segment, C, carrying out recovery, conversion, sequencing and sequence analysis processes on PCR products, D, carrying out 5 ' RACE and 3 ' RACE processes and splicing a sequencing result to obtain a full-length cDNA sequence, and E, designing specific primers in 5 ' UTR and 3 ' UTR zones of the full-length cDNA sequence according to the full-length cDNA sequence to obtain a molecular marker. The invention also discloses an application of a restoring gene molecular marker in hybrid rape parent breeding. The preparation method has the advantages of improvements of a speed of restorer seeding and an efficiency of verification, simple and easy method, good operability, rapid identification speed of restoring genes, great reduction of necessary workload of test cross and progeny planting observation in a restorative verification process, and improvement of an efficiency of hybrid seeding.
Owner:INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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