31results about How to "No sequencing required" patented technology

The invention relates to an authenticating method of fish hybrids and particularly relates to an RAG2 molecular marker RFLP identifying method for aquaculture of catfish hybrids. Southern catfishes, catfishes and hybrids of the southern catfishes and the catfishes serve as objects of study, RAG2 gene sections in catfishes are selected to serve as molecular markers, the RFLP analysis of the RAG2 gene sections is achieved, and the southern catfishes, the catfishes and hybrids of the southern catfishes and the catfishes are identified successfully. The method has the advantages of being fast in experiment, convenient to operate, direct in identification and free from sequencing.

The invention relates to a kit for simultaneous detection of Anopheles moltatus, Anopheles reckeii and Anopheles lanceolata by multiplex PCR. The kit includes specific primers for Anopheles papillonus, Anopheles reckeii, and Anopheles beriorhynchus, 10×buffer, dNTPs, MgCl 2 , Taq enzyme, template DNA, sterile double-distilled water, the sequences of the specific primers of Anopheles moltatus, Anopheles reckeii, and Anopheles lanceolata are respectively as SEQ ID NO.5-6, SEQ ID NO.15 -16, shown in SEQ ID NO.33-34. The multiplex PCR detection kit of the present invention can give qualitative detection results within 3 hours after receiving the DNA sample, saves cost and time, and has low requirements for personnel and equipment. and an effective tool for An.

The invention relates to a related specie identification technology, in particular to a method for identifying related species of an oyster. The method specifically comprises the following steps of: by using an oyster genome DNA (Deoxyribonucleic Acid) as a template, designing a primer according to a tag sequence of the oyster specie genome; carrying out PCR (Polymerase Chain Reaction) amplification on the primer; and distinguishing oyster species by using Tm value difference in a high-resolution melting curve of an amplification product. Through the adoption of the method provided by the invention, the species can be identified rapidly, economically, simply and conveniently; and compared with a tag sequence sequencing result, an identifying result of the method provided by the invention is 100% in accuracy.

The invention provides a primer-free gene synthesis method based on a plasmid library, and the method is as follows: constructing a plasmid library containing 16384 plasmids (any combination sequence of four basic groups of 47 and 7bp), starting from a constructed plasmid containing 7 bp, dividing a selected target gene according to each 82bp, successively reconstructing four resistance genes Kana, cam, gem and spc by enzyme digestion and enzyme connection operation to obtain a plurality of 82bp plasmids, and assembling the 82bp plasmids into long fragment genes. According to the method, after construction of the plasmid library containing the 16384 plasmids (any combination sequence of four basic groups of 47 and 7bp), various target genes can be synthesized without any primer. The synthesis method has the advantages of simple operation, extremely low mutation rate, no need of sequencing, low cost and less amount of screening, is conductive to constructing a mutant library, can realize solution operation, and is easy to realize automation.

The invention relates to an ITS2 molecular marker identifying method for aquaculture of catfish hybrids. Southern catfishes, catfishes and hybrids of the southern catfishes and the catfishes serve as objects of study, ITS2 gene sections in catfishes are selected to serve as molecular markers, the length difference the IST2 gene sections of the southern catfishes and the catfishes, and the southern catfishes, the catfishes and hybrids of the southern catfishes and the catfishes are identified successfully. The method has the advantages of being fast in experiment, convenient to operate, visual in identification and free from sequencing.

The invention relates to a group of specific primers and a method for identifying stiff silkworms, belonging to the technical field of identification of traditional Chinese medicines. According to themethod, after a stiff silkworm sample is crushed, the deoxyribonucleic acid (DNA) in domestic silkworms and the DNA in fungi are extracted at the same time by using a CTAB method; the extracted DNA is taken as a template, specific nucleotide sequences, which are designed and screened, are used as primers for performing polymerase chain reaction (PCR) amplification, and the products are then subjected to agarose gel electrophoresis analysis and gel imaging system detection, so that the authenticity and adulteration of the stiff silkworms can be finally detected based on the detection results.The method provided by the invention can be used for judging whether the sample is a stiff silkworm fake product or adulterated product, is simple to operate, does not needing sequencing, is high in specificity, rapid and accurate, and has unique advantages and important application value in the authenticity identification of stiff silkworm samples.

The invention relates to a kit for simultaneously detecting Anopheles bechai, Anopheles multicolored and Anopheles aconitum by multiplex PCR. The kit includes specific primers for Anopheles glabra, Anopheles multicolored, and Anopheles aconitum, 10×buffer, dNTPs, MgCl 2 , Taq enzyme, template DNA, sterile double-distilled water, the sequences of the specific primers of Anopheles mechai, Anopheles multicolored, and Anopheles aconitum are respectively as SEQ ID NO.5-6, SEQ ID NO.13 -14, shown in SEQ ID NO.25-26. The multiplex PCR detection kit of the present invention can give qualitative detection results within 3 hours after receiving the DNA sample, saves cost and time, and has low requirements for personnel and equipment. and an effective tool for Anopheles aconitum.

The invention relates to an improved primer for detecting the dynamic mutation of a CAG repetitive sequence of a spinocerebellar ataxia ATXN3 gene and a method thereof, characterized in that a specially designed specific primer is adopted to amplify the amount of a DNA fragment containing the CAG repetitive sequence in the ATXN3 gene to the observable degree through a special PCR procedure, and gel electrophoresis is utilized to observe the size of a PCR amplification product fragment and estimate the number of replication of the CAG dynamic mutation in the ATXN3 gene. The invention has the advantages that: 1. simplicity and economy, and no sequence detection, i.e. the method can be operated in common molecular biology laboratories or control laboratories, thereby establishing a simple detection method of the amplification CAG repetitive sequence; 2. high sensitivity and reliability: 25ng genome DNA is enough to detect the number of replication of the CAG dynamic mutation in the ATXN3gene, thereby a reliable basis is provided for clinically diagnosing spinocerebellar ataxia.