34results about How to "Guaranteed amplification efficiency" patented technology

The invention relates to a nucleic acid releasing agent, a nucleic acid PCR (polymerase chain reaction) amplification method and a PCR amplification kit. The nucleic acid releasing agent comprises Tris-HCl, sodium chloride, potassium chloride, Tween 20, Triton X-100, Nonidet P-40 and strong base; the molar concentration of the Tris-HCl is 0.5-500mM, the molar concentration of the sodium chloride is 20-500mM, the volume percentage of the Tween 20 is 0.1-2%, the volume percentage of the Triton X-100 is 0.1-3%, the volume percentage of the Nonidet P-40 is 0.1-3%, the mass concentration of the potassium chloride is 5-8mg / mL, and the mass concentration of the strong base is 2-50mg / mL. The nucleic acid releasing agent has the advantages that RNA-containing samples can directly release RNAs at the room temperature, and can be directly mixed with PCR reaction liquid to achieve PCR amplification, so that amplification can be completed without complex nucleic acid extraction and purification processes.

The invention belongs to the technical field of PCR (polymerase chain reaction), and particularly relates to a digital PCR chip with silicon substrate arrays and micro-reaction pools and a method for manufacturing the digital PCR chip. The digital PCR chip mainly comprises an upper cover and a chip. Micro-pores which are arrayed to form honeycombs are formed in the chip, and the upper cover is fixed onto a chip groove. The method includes particular steps of selecting a silicon chip with a single polished surface; cleaning the silicon chip; coating a layer of uniform photoresist on the polished surface of the silicon chip in a spinning manner; forming circular graphic arrays by means of exposure; etching silicon by the aid of a dry process in masks, namely, the circular graphic arrays to form micro-pore structures; removing the photoresist and scribing the chip to completely manufacture the digital PCR chip.

The invention relates to a nucleic acid composition for detecting hereditary anemia, a detection kit and a use method thereof. The nucleic acid composition and detection kit for the detection of hereditary anemia can be used for the simultaneous detection of 4 deletion-type α-thalassemia genes, 3 non-deletion-type α-thalassemia genes, 19 mutant-type β-thalassemia genes, and sickle-shaped anemia Genetics such as anemia and specific types of genetic mutations. Compared with the existing similar technologies, the hereditary anemia detection kit has added several mutation detection types of hereditary anemia, such as α THAI Deletion thalassemia, sickle cell anemia, and two less common types of beta-thalassemia, the 71 / 72(+T) mutation and the ‑28M(A‑C) mutation. The increased detection of these types of hereditary anemia can provide intuitive reference and prompts for clinical detection of hereditary anemia, thus greatly reducing the risk of missed detection of clinical hereditary anemia and reducing the birth rate of children with severe anemia.

The invention belongs to the field of genetic engineering and in particular relates to novel endoglucanase, coding genes cel5A-h31 and application thereof. A nucleotide sequence of genes cel5A-h31 coding the endoglucanase is shown as SEQ ID No. 1. Bioinformatics analysis shows that the endoglucanase coded by the genes belongs to the fifth family. The endoglucanase is successfully cloned by the inventor by virtue of a design primer and successfully expressed in a prokaryotic expression system, the recombinant endoglucanase obtained by induced ultrasonic purification has an optimal operating pHvalue of 5.0 and an optimal temperature of 55 DEG C, can maintain relatively stable enzymatic activity at a temperature of 50 DEG C or below, has high pH tolerance and can maintain the enzymatic activity of 75% or higher at the pH value of 5.0-9.0. The recombinant endoglucanase has high research and industrial application potential.

The invention relates to the field of genetic engineering and in particular relates to endoglucanase as well as an encoding gene cel5A-h42 and application thereof. The gene cel5A-h42 for encoding theendoglucanase has a nucleotide sequence shown as SEQ ID No. 1. Bioinformatic analysis shows that the endoglucanase encoded by the gene belongs to the fifth family of glycoside hydrolase. The inventorsuccessfully clones the encoding gene through a designed primer and the encoding gene is successfully expressed in a prokaryotic expression system; the recombinant endoglucanase obtained by inducing ultrasonic purification has the optimum effect pH (Potential of Hydrogen) of 5.0 and the optimum temperature of 55 DEG C, and can keep relatively stable enzyme activity at 50 DEG C. The enzyme has relatively strong pH tolerance when the pH is 5.0 to 10.0 and 70 percent or more of the enzyme activity can be kept. The recombinant endoglucanase has a relatively great research and industrial application potential.

The invention discloses sugarcane genome endogenous reference gene isopentenyl diphosphate delta-isomerase gene PCR (polymerase chain reaction) primer sequences and an amplification method. The primer sequences are composed of ScIDI2-F and ScIDI2-R. The PCR amplification method comprises a reaction system and amplification conditions. The invention provides a genome endogenous reference gene for PCR detection and species specific identification of transgenic sugarcane, and meanwhile, provides insurance for detection precision and result accuracy of the transgenic sugarcane.

The invention relates to the genetic engineering field, and concretely relates to a whole cell xylanase for promoting rumen fermentation, and a preparation method thereof. The whole cell xylanase for promoting rumen fermentation is characterized in that a gene cloning and yeast surface display technology is used to display a xylanase coding gene ORF6-UN from a Hu sheep rumen microorganism Fosmid library on the surface of Saccharomyces cerevisiae, and inducible expression, centrifuging, cell precipitate recovery and lyophilization are carried out to obtain dry cell powder which is ruminal whole cell xylanase; and the nucleotide sequence of the xylanase coding gene ORF6-UN is represented by SEQIDNo.1. The whole cell xylanase for promoting rumen fermentation can be supplemented to animal diets as a feed additive, and can increase the nutrition intake of animals and improve the fibrous substance utilization efficiency of the animals.

The invention relates to the field of genetic engineering and in particular relates to endoglucanase as well as an encoding gene cel5A-h37 and application thereof. The gene cel5A-h37 for encoding theendoglucanase has a nucleotide sequence shown as SEQ ID No. 1. Bioinformatic analysis shows that the endoglucanase encoded by the gene belongs to the fifth family of glycoside hydrolase. The inventorsuccessfully clones the encoding gene through a designed primer and the encoding gene is successfully expressed in a prokaryotic expression system; the recombinant endoglucanase obtained by inducing ultrasonic purification has the optimum effect pH (Potential of Hydrogen) of 6.0 and the optimum temperature of 40 DEG C, and can keep relatively stable enzyme activity at 40 DEG C. The enzyme has relatively strong pH tolerance when the pH is 5.0 to 8.0 and 80 percent or more of the enzyme activity can be kept. The recombinant endoglucanase has a relatively great research and industrial applicationpotential.

The invention belongs to the field of genetic engineering and particularly relates to an endoglucanase and its coding gene cel5A-h38 and application. The gene cel5A-h38 for encoding the endoglucanasehas a nucleotide sequence shown in the formula of SEQ ID No. 1. The bioinformatic analysis result shows that the endoglucanase coded by the gene belongs to the 5th family of glycoside hydrolase. Primers are designed and cloned and then are successfully expressed in a prokaryotic expression system. The recombinant endoglucanase obtained by induced ultrasonic purification has the optimal pH of 6.0 and the optimal temperature of 45 DEG C, keeps the stable enzyme activity at 50 DEG C or less, has strong pH tolerance and keeps the enzyme activity of 60% or more at pH of 5.0-10.0. The recombinant endoglucanase has the large research and industrial application potential.