60results about How to "Improve the efficiency of database construction" patented technology

Based on a test platform of the Illumina Company at present and aiming at the characteristic of a small section of a Fosmid library, a plurality of libraries are mixed together to form one library; furthermore, a purifying step is omitted, so a new Fosmid library preparation flow is established. The Fosmid library is constructed successfully; the Fosmid library is successfully used for sequencing; therefore, the library construction time is saved and the sequencing cost is reduced.

The invention belongs to the technical field of gene engineering, and particularly relates to a detection method of a low frequency mutated DNA fragment and a library building method. The technical problems that in the existing detection technique, the DNA fragment capturing efficiency is low, wrong mutation is introduced in the library building process, the detection limit is low, and the sensitivity is low are solved by adopting the hybrid capturing and molecular tagging technique and meanwhile combining with a special library building mode, the library specificity is improved, the false positive rate is lowered, and the template utilization rate is improved.

The invention discloses a rapid library building method for identifying a target protein chromatin binding map, which comprises the following steps: collecting sample cells, adding activated magneticbeads into the cells, incubating, and incubating the magnetic beads with a primary antibody; pre-binding a secondary antibody with recombinant fusion transposase; incubating the magnetic beads with the pre-combined secondary antibody and recombinant fusion transposase, adding a transposase activator and a cell perforating agent into the magnetic beads, and incubating; and terminating the transposase reaction, and carrying out library amplification. According to the method, the process of a traditional method is remarkably simplified, the library building time is shortened, the library buildingefficiency is improved, the loss is small, and the requirement for library building materials is lower. Meanwhile, three DNA standard substances with different concentrations are doped into the library building process, so that the effect of accurately quantifying the DNA copy number in the library can be achieved. The method is very suitable for researching a small amount of precious samples orsamples with different treatment states.

The invention provides a construction method of a cell Hi-C sequencing library. The construction method comprises the following steps: carrying out formaldehyde cross-linking immobilization on 10<5>-order cells by adopting a cross-linking system of which the volume is less than or equal to 800 muL and the final concentration of formaldehyde is 1.5-2.2%, and then treating the cross-linked cells byadopting an optimized cracking system and temperature to obtain a cell nucleus suspension; carrying out Hi-C library construction on the cell nucleus to obtain an Hi-C sequencing library. The cell membrane lysis solution comprises the following components with final concentrations: Tris-HCl with the pH value of 7.8 to 8.2 and the concentration of 9 to 11mM, NaCl with the concentration of 9 to 11mM, CA-630 with the concentration of 0.1 to 0.3 percent and a protease inhibitor with the concentration of 1*. A small-volume system fixes the initial quantity of low cells, optimizes a cell membrane lysis system and reaction conditions, ensures the integrity of cell nucleuses, and improves the quantity and purity of DNA of the cell nucleuses, thereby improving the library building efficiency (improving the proportion of effective data).

The invention relates to an intelligent question answering knowledge base establishment method, establishment device and establishment system. The establishment method comprises the following steps: providing a domain knowledge data base, wherein the domain knowledge data base comprises a plurality of pieces of preset knowledge; receiving initial request information; performing semantic similarity calculation on the initial request information and the preset knowledge in the domain knowledge data base, and judging whether the maximum value of the semantic similarity calculation results is larger than a similarity threshold or not; if the maximum value of the semantic similarity calculation results is larger than the similarity threshold, storing a standard question and an extended question in the preset knowledge corresponding to the initial request information and the maximum value of the semantic similarity calculation results in the intelligent question answering knowledge base; if the maximum value of the semantic similarity calculation results is smaller than the similarity threshold, carrying out the abstract semantic recommendation process so as to acquire one or more specific semantic expressions corresponding to the initial request information, and storing the initial request information and the specific semantic expressions in the intelligent question answering knowledge base. The establishment method provided by the invention can improve the establishment efficiency of the intelligent question answering knowledge base.

The invention discloses a kit for DNA library building and an application of the kit, and in particular relates to a kit for peripheral blood cfDNA library building and an application of the kit. According to the invention, the kit for DNA library building is firstly provided, wherein the kit comprises a component A and a component B; the component A consists of the following ingredients at a ratio as follows: 1-3U of T4 PNK to 1-4.5U of Klenow Fragment to 0.25-0.5U of LA Taq DNA Polymerase to 0.3-0.9U of T4 DNA Polymerase to 0.5-2.5 nmol of dNTP to 0.5-2.5nmol of dATP; and the component B consists of the following ingredients at a ratio as follows: 2.5-8nmol of ATP to 26.5 132U of T4 DNA Ligase to 0.213 0.534[mu]L of PEG 8000. According to the kit and the application method provided by the invention, DNA loss in a library building process can be reduced to the greatest extent, the usage amount of plasma can be reduced, library building cost can be reduced, library building time can beshortened, operations can be simplified and more stable and reliable results can be obtained. The kit provided by the invention is significant for improving library building efficiency and reducing labor cost.

According to the invention, wild type DNase I related sites are mutated and screened to obtain three mutants with good effects, wherein two mutants are in double-site mutation, and mutation sites are respectively A249T, R42H and A249T, F110V; one mutant is in three-site mutation, and the mutation sites are A249T, R42H and F110V. The DNase I mutant with the three-site mutation has the best effect, and the DNase I mutant has an amino acid sequence shown in SEQ ID No.1 and a coding nucleotide sequence shown in SEQ ID No.2. The invention also discloses application of the DNase I mutants in construction of a high-throughput sequencing library. The DNase I mutants disclosed by the invention can efficiently and stably break genome DNA, fragmented products of the DNase I mutants can be applied to construction of a high-throughput sequencing library, and compared with wild type DNase I, library construction and sequencing quality is more excellent, and GC preference is lower. Low-cost and high-efficiency library building can be realized, and sequencing quality is excellent.

The invention provides an mRNA library construction method. The mRNA library construction method includes mixing an mRNA fragment with a first mixed enzyme solution and a first buffer solution to obtain a first reaction system, and subjecting the first reaction system to first heat treatment to obtain a first reaction solution; mixing the first reaction solution with a second mixed enzyme solutionand a second buffer solution to obtain a second reaction system, and subjecting the second reaction system to second heat treatment to obtain a second reaction solution; mixing the second reaction solution with T4DNA ligase, a third buffer solution and a Y-shaped joint to obtain a third reaction system, subjecting the third reaction system to third heat treatment, and after the third heat treatment, performing purification to obtain a third reaction solution; adding a pre-mixed solution and a amplification primer into the third reaction solution to obtain a fourth reaction system, subjectingthe fourth reaction system to fourth heat treatment, and after the fourth heat treatment, performing purification to obtain an mRNA library. The mRNA library construction method and a kit, which are beneficial to improvement of the library construction efficiency, can be provided.

The invention relates to the fields of genetic engineering and molecular biology, and provides a rapid library building method, an SNP (single nucleotide polymorphism) genotyping method and a sequencing method. The rapid library building method includes the following steps: A. a sample to be sequenced goes through PCR (polymerase chain reaction) amplification, so as to obtain an amplified product; during the PCR amplification process, at least one PCR primer in a PCR primer pair for PCR amplification contains a breakable site or a cleavable sequence; B. the amplified product is added to a cleavage-ligation reaction system for cleavage and ligation, so as to obtain a library molecule containing a first linker; the cleavage-ligation reaction system comprises a ligase, a clastogen, the first linker and a ligation buffer solution; the clastogen is used for specific cleavage of the breakable site or the cleavable sequence in the amplified product, so as to form a first cohesive end; the first linker is a nucleic acid molecule, and contains a second cohesive end capable of completely complementary pairing with the first cohesive end. The invention simplifies experiment steps, and improves the library building efficiency.

The invention belongs to the fields of genetic engineering and molecular biology, and in particular relates to a method for constructing a high-throughput sequencing library. The steps include (1) DNA fragmentation; (2) purification; (3) linker connection; (4) amplification; (5) small fragment removal; (6) library fixation and other steps. The joint connection efficiency of the present invention is high, and by using the method of the present invention, the sequencing plate can be used simply and efficiently, the loss of samples and reagents is reduced, and a high-throughput sequencing library can be constructed.

The invention discloses mutant type Taq enzyme capable of improving the A adding efficiency as well as a preparation method and application of the mutant type Taq enzyme. The preparation method comprises the following specific steps: directionally mutating wild type Taq enzyme, and mutating 315th glutamic acid into lysine; mutating 388th glutamic acid into valine, mutating 507th glutamic acid into the lysine, mutating 578th aspartic acid into glycine, mutating 608th alanine into the valine, and mutating 747th methionine into arginine to obtain the mutant type Taq enzyme with the amino acid sequence shown as SEQ NO.1. The A adding efficiency of the mutant type Taq enzyme is improved, and the database building efficiency is improved; the mutant type Taq enzyme can be applied to a template with lower initial amount, and the completeness and the cover degree of building the database are improved.