74 results about "Pcr sequencing" patented technology

The invention relates to the technical field of bioinformatics, particularly to a whole-genome methylation non-bisulfite sequencing library, a construction and applications thereof, wherein the sequencing library comprises a TET enzyme reaction solution, the TET enzyme reaction solution comprises the following independently packaged components: a TET enzyme oxidation buffer solution, and the TET enzyme oxidation buffer solution comprises, by micromole, (20-167)*10<3> parts of HEPES or Tris-Cl, (100-333)*10<3> parts of NaCl, 3.3*10<3> parts of alpha-KG or 2-oxoglutarate, 6.67*10<3> parts of ascorbic acid and 4*10<3> parts of adenosine triphosphate. According to the invention, by combining the kit, Fe(NH4)2(SO4)2 and TET enzyme, 5mc can be oxidized into 5cac, the 5cac is reduced into dihydrouracil under the action of a reducing agent, and T is identified through PCR sequencing, so that the the DNA methylation C-to-T conversion under the non-bisulfite condition is achieved, the defects ofbase imbalance and low sequencing data use rate of the existing methylation sequencing library constructed based on the bisulfite conversion are solved; and the formula further has effects of simplecomponents, extremely low TET enzyme use amount and significant cost reducing.

The invention provides a kit used for staphylococcus aureus detection. The kit used for staphylococcus aureus detection contains guide RNA of a specific targeted staphylococcus aureus rpoB gene, an amplification primer pair, hydrated TwistAmp basic kit reaction drying balls, LbCas12a protein, a single-stranded DNA probe (ssDNA), Ribonuclease Inhibitor and buffer liquid. By using the kit for staphylococcus aureus detection, the detection specificity is high, and staphylococcus aureus and other staphylococcus, including staphylococcus epidermidis, staphylococcus hominis, staphylococcus warneri,staphylococcus capitis and staphylococcus haemolyticus, can be separated. Meanwhile, by using a RNA sequence for detection, elapsed time is about 1 hour, no PCR instruments are used, the equipment requirements are low, and are significantly lower than that of a traditional bacterial culture method or PCR sequencing method, and the clinical application valve is large.

This present invention identifies mutations in several androgen-metabolic genes (SRD5A2, CYP17, HSD3B2, and HSD17B3) and methods of using such mutations in the diagnosis and treatment of inheritable prostate cancer susceptibility. Isolation of genomic DNA of various racial / ethnic populations followed by SSCP scanning and direct PCR sequencing of the aberrant SSCP (single-strand conformation dependent DNA polymorphism) patterns allows for identification of the disclosed polymorphisms. Screening for the disclosed mutations establishes a differential distribution among various racial / ethnic groups as well as altered in vivo enzyme activity that parallels prostate cancer risk.

The invention discloses a method for evaluating genetic relationship DNA (deoxyribonucleic acid) molecules of a mulberry variety. The method comprises the steps of (1) system development analysis of the mulberry variety: 1) material selection; 2) screening of a contrast sequence; 3) extraction of mulberry genome DNA and detection of purity and concentration; 4) PCR (polymerase chain reaction) amplification; 5) PCR detection; 6) PCR sequencing; 7) sequence splicing and comparison, and identification of an evolutionary relationship; (2) genetic relationship analysis of the mulberry variety. According to the method disclosed by the invention, the system development analysis is firstly performed, materials are reasonably selected from worldwide mulberry varieties and variants, and then genetic diversity analysis of mulberry variety populations is performed to further determine the genetic relationship, so that the accuracy and repeatability of evaluating the genetic relationship are significantly improved by using the two-step analysis method, and a great promotion effect is achieved for protection of mulberry variety resources, mulberry variety fingerprint identification, molecular aided breeding and variety resource development and utilization.

The invention provides an optimized separation method for human intestine probiotics. The optimized separation method comprises the following steps that 1 basic medium preparing, wherein a mucoprotein liquid medium and a mucroprotein solid medium are prepared; 2 screening, wherein a feces specimen is collected and inoculated in the mucroprotein liquid medium for anaerobic culturing to obtain bacterium liquid, the bacterium liquid is taken as a template, a PCR experiment is performed, the bacterium liquid in a PCR positive reaction tube is selected to be inoculated in the mucroprotein solid medium for anaerobic culturing to obtain a bacterium colony, the bacterium colony on the mucroprotein solid medium is inoculated onto a chocolate plate for anaerobic culturing to obtain a bacterium colony, the bacterium colony on the chocolate plate is inoculated onto a Columbia blood plate for purification culturing to obtain a bacterium colony, and PCR sequencing verification is performed on the bacterium colony on the Columbia blood plate through a 16sRNA universal primer to separate the human intestine probiotics. The optimized separation method for the human intestine probiotics is practical, simple, visualized and convenient.

The invention provides a kit for detecting staphylococcus haemolyticus. The kit comprises guide RNA specifically targeting staphylococcus haemolyticus rpoB gene, an amplification primer pair, hydration TwistAmp basic kit reaction drying balls, LbCas12a protein, a single-chain DNA probe (ssDNA), a Ribonuclease Inhibitor and a buffer solution. When the kit is used for detecting the staphylococcus haemolyticus, high detection specificity is achieved, and the staphylococcus haemolyticus can be distinguished from other staphylococcus such as staphylococcus aureus, staphylococcus hominis, staphylococcus warneri, staphylococcus capitis and staphylococcus epidermidis. Meanwhile, when the RNA sequence is used to perform detection, detection time is about 1 hour, a PCR instrument is not needed, equipment requirements are evidently lower than those of a traditional bacterial culture method or a PCR sequencing method, and a high clinical practical value is achieved.

The invention discloses a linearized DNA vector pHB-1 plasmid and a test kit prepared from the same. The genetic sequence of the linearized DNA vector pHB-1 plasmid is shown as SEQ ID NO: 1; the testkit comprises a linearized DNA vector pHB-1 plasmid, an efficient T4DNA ligase, a PCR sequencing primer tL3-seq and an experimental control substance. Wherein the experimental control comprises a specific gRNA sequence primer for editing an escherichia coli MlaC gene, a DNA repair template for introducing MlaC L11R point mutation and a PCR sequencing primer for confirming the MlaC L11R point mutation; the test kit disclosed by the invention can be used for carrying out gene editing experiments on various bacteria, has repeatability and stability, and can be used for editing one or more targetgenes, and the editing efficiency can reach 90-100%.

The present invention provides a kit for detecting staphylococcus warneri. The kit comprises a guide RNA specifically targeting a staphylococcus warneri rpoB gene, an amplification primer pair, hydration TwistAmp basic kit reaction drying spheres, LbCas12a protein, a single-stranded DNA probe (ssDNA), an ribonuclease inhibitor and a buffer solution. The kit is used for detecting the staphylococcuswarneri and high in detection specificity, and can separate the staphylococcus warneri from other staphylococci, including staphylococcus aureus, staphylococcus hominis, staphylococcus epidermidis, staphylococcus capitis and staphylococcus haemolyticus. At the same time, the use of the RNA sequence for the detection takes about 1 hour, does not require a PCR instrument, has low requirements on equipment, and is significantly lower than a conventional bacterial culture method or a PCR sequencing method and large in clinical practical value.