317 results about "SYBR Green I" patented technology

The invention relates to a real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and a detection method thereof. The sequences of the primer are described as follows: an upstream primer P1: 5'-CAGATCCCCGGGCACGAGA-3'; a downstream primer P2: 5'-CCTCACCTTCAGGGATTC-3'. The detection method includes the steps of: establishing the real-time fluorescence quantification PCR method based on SYBR Green I with the primer, wherein difference of temperature of a melting curve exists due to the difference of GC content of a nucleotide in a specific gene fragment region of gene 1-type duck circovirus and gene 2-type duck circovirus which are amplified by the primer, so that infection of the gene 1-type duck circovirus and the gene 2-type duck circovirus can be directly carried out according to the difference of Tm value in the melting curve. The method can distinguish the infection caused by the duck circovirus in different genotypes just with one group of primer, thereby providing technical fundament for healthy breeding in duck breeding industry.

The invention discloses a method and a primer composition for detecting soybean fusarium oxysporum based on an LAMP (loop-mediated isothermal amplification) technology. The primer composition comprises four specific primers FIP, BIP, F3 and B3 for detecting the LAMP molecules of the soybean fusarium oxysporum, and two ring primers LF and LB used for increasing the reaction speed. The method is used for performing LAMP on DNAs to be detected by the primer composition provided by the invention; a detection result is observed by naked eyes or under the irradiation of ultraviolet light with the wavelength of 245 nm; the color change and the fluorescence intensity of SYBR Green I are taken as result judgment standards. The invention provides the new molecular detection method and the primer composition for detecting the soybean fusarium oxysporum, the soybean fusarium oxysporum is subjected to LAMP detection, the detection cycle is short, the accuracy is high, the sensitivity is high, and the detection result is observed by the naked eyes.

The invention discloses a multiplex SYBR Green I real-time fluorescence PCR (polymerase chain reaction) detection primer and method for porcine rabies virus, porcine parvovirus and porcine circovirus type 2. The primer is obtained through synthesis according to design. The multiplex SYBR Green I real-time fluorescence PCR detection method for detecting porcine rabies virus, porcine parvovirus and porcine circovirus type 2 by utilizing the primer comprises the following steps: extracting the DNA of a sample, and then, detecting the sample by utilizing a SYBR Green I real-time fluorescence PCR reaction system and a SYBR Green I real-time fluorescence PCR amplification program. The invention has the beneficial effects that three types of viruses, namely the porcine rabies virus, the porcine parvovirus and the porcine circovirus type 2, can be simultaneously and effectively diagnosed and detected; non-specific swine fever virus, porcine reproductive and respiratory syndrome virus and swine influenza virus can not be detected; and the invention is beneficial to identification and diagnosis of the breeding disorder virus of a pregnant swine, and has better sensitivity, repeatability and stability.

The invention belongs to the technical field of detection of red-sea bream iridovirus infection, and relates to a real-time quantitative detection method of polymerase chain reaction (PCR) technology by adopting a fluorescent probe or an SYBR GREEN I fluorescent dye, in particular to a real-time quantitative PCR detection method for red-sea bream iridovirus. The method comprises the following steps of: designing a specific primer sequence and a fluorescent probe sequence, then establishing a real-time quantitative PCR system and a reaction program, preparing plasmids containing a target amplification fragment as a standard substance to draw a standard curve, and finally establishing a result judgment base of a sample to be detected. The method has the advantages of simple detection, high detection speed, good effect, accurate quantification, strong specificity, high sensitivity and the like.

The invention discloses a prawn IHHNV (infectious hypodermal and hematopoietic necrosis virus) nucleic acid isothermal amplification detection kit which comprises a grinding fluid tube containing a grinding fluid, a nucleic acid extracting solution tube A containing a sodium acetate solution, a nucleic acid extracting solution tube B containing absolute ethyl alcohol, a nucleic acid extracting solution tube C containing an alcohol solution with a mass concentration of 70 percent, a TE (tellurium) buffer solution tube containing a TE buffer solution, a UNG (uracil-DNA glycosidase) enzyme tube containing uracil DNA (deoxyribonucleic acid) glycosylase, an LAMP (Loop-mediated isothermal amplification) reaction liquid tube containing an LAMP reaction liquid, a BstDNA polymerase tube containing Bst DNA polymerase, a color-developing agent tube containing nucleic acid dye SYBR Green I, a positive control nucleic acid tube containing prawn IHHNV positive DNA and a negative control tube containing sterilizing double distilled water. The invention also provides a method for detecting the prawn IHHNV by utilizing the detection kit. The method has the characteristics of low cost, high detection sensitivity and easy site operation.

The invention relates to a method for detecting the expression quantity of pathogenic genes of rice blast germs in infected rice leaf tissue. The invention provides the method for detecting the expression quantity of the pathogenic genes in different time points after rice is infected by the rice blast germs. The method adopts a technical proposal that the rice blast germs are inoculated on a rice leaf by spraying, sampling is performed and total RNA is extracted; real-time fluorescent quantitative PCR detection is performed on pathogenic genetic fragments of rice blast germs by an SYBR Green I embedded fluorescence method; the C(t) value of a sample to be detected is recorded; and the expression quantity of the related pathogenic genetic fragments is calculated according to the formula. The method can quickly and accurately detect the expression quantity of the pathogenic genes of the rice blast germs on different periods of time in the initial stage of inoculation of the rice blast germs, analyzes the expression quantity of different pathogenic genes in different rice cultivars and the variation of the different pathogenic genes on different periods of time after inoculation, and analyzes the spatiotemporal expression mode of the related pathogenic genes of the rice blast germs and so on. The method has accurate detection result and good repeatability, and is favorable for explaining the functions of the genes.

The invention discloses a loop isothermal amplification primer for quickly detecting tylenchulus semipenetrans and application of the loop isothermal amplification primer. The loop isothermal amplification primer comprises a primer pair TSF3 / TSB3 and a primer pair TSFIP / TSBIP, and the sequences of the primer pairs are shown as SEQ ID NO. 1 to 4 respectively. A loop isothermal amplification method is established through the loop isothermal amplification primer, loop isothermal amplification is performed through taking a sample DNA as a template, and results can be judged in two ways after the end of reaction, wherein the first way is that an amplification primer is subjected to electrophoresis, and a sample of a specific ladder-shaped strip, which occurs, is judged to be positive; the second way is that through adding SYBR green I into a reaction system, a sample of which the color is changed is observed with naked eyes to be positive. The loop isothermal amplification method is low in requirement for instruments and equipment, quick, safe, and high in specificity and sensitivity; a technical support is provided for the detection of the tylenchulus semipenetrans, in particular to the quick detection work of a grass-root quarantine unit for the tylenchulus semipenetrans, and the popularization and application values are very high.