262 results about "Dna concentration" patented technology

The present invention relates to methods for detecting free floating nucleic acids, as present in not cellular bound nucleic acids in bodily fluids like plasma or serum fractions of human or animal blood or in any other tissue samples derived from the human or animal body in order to diagnose a cell proliferative disease. Specifically the invention relates to the detection of increased levels of nucleic acids in bodily fluids. Furthermore the invention allows to determine the source of the enriched DNA by measuring the ratio of DNA originating from a certain organ versus total DNA from other organs in a given bodily fluid sample by specifying the DNA's methylation pattern. This can be done with or without increasing the DNA concentration of a given biological sample. In a preferred embodiment a further analysis of this methylation pattern allows for the detection of the presence of tumourous or otherwise proliferative disease in said organ.

The invention discloses an HBV (hepatitis B virus) fluorescent quantitative PCR (polymerase chain reaction) detection kit which comprises a probe for detecting the target polynucleotide, an upstream primer and a downstream primer for amplifying the target polynucleotide, a PCR buffer solution and DNA (deoxyribonucleic acid) polymerase, wherein the probe for detecting the target polynucleotide hasa SEQ No.1 sequence. As the probe for detecting the target polynucleotide has the SEQ No.1 sequence, the kit disclosed by the invention has good specificity; by using the kit provided by the invention, the HBV-DNA concentration in the unknown sample such as serum, plasma or milk can be quickly and accurately measured; and the kit has good specificity and can detect eight gene types of HBV.

The invention discloses a fluorescence quantitative PCR detection kit of hepatitis B virus and an application thereof. The kit is composed of the following independent components: DNA extraction solution I, DNA extraction solution II, DNA extraction solution III, DNA extraction solution IV, positive control interior label, PCR reaction liquid, probe HBV-SP, enzyme mixed liquor containing heat resistant DNA polyase and uracil DNA glycosylase, quantitative hepatitis B virus reference material, hepatitis B virus positive control serum and hepatitis B virus negative control serum, wherein DNA extraction solution I contains 0.2-1.0% of lauryl sodium sulphate (mass/volume), 1.0-4.0% of Triton (volume/volume) and 0.2-1.0mol/L of guanidinium isothiocyanate; DNA extraction solution II contains 100-300mmol/L of 4-HEPES, 100-300mmol/L of sodium chloride with pH of 6.5+/-0.2 and 100-400 mu g/ml of magnetic beads; DNA extraction solution III contains 0.1-1.0% of Triton (volume/volume) and 100-300mmol/L of sodium chloride; DNA extraction solution IV contains mineral oil. The fluorescence quantitative PCR detection kit of hepatitis B virus of the invention can be used for detecting the HBV-DNA concentration in samples of serum, blood plasma or latex and the like.

Disclosed are systems and methods for identifying and quantitating the presence of one or more DNA species in a sample population through PCR amplification. DNA species quantitation includes a determination of a threshold fluorescence value used in the assessment of the PCR amplification reaction. Various embodiments of the present invention incorporate an enhancement function useful in selecting appropriate threshold fluorescence values and facilitate the determination of DNA concentrations by quantitative PCR based methodologies.

The invention relates to the microbiology and molecular biology technical field, in particular to a method for extracting pig manure sample total DNA. The method mainly comprises manure sample pretreatment and sample total DNA extraction. Moreover, the method comprises the following steps that: collected fresh manure sample or frozen manure sample is taken as a material; the pretreatment of the manure sample is carried out by means of a PBS buffer solution, a paraformaldehyde solution and anhydrous alcohol so as to obtain manure sample bacteria suspension; and a CTAB solution and beta-mercaptoethanol, etc. are used to carry out cell fragmentation, thereby releasing DNA. The method has the characteristics of simple operation, quickness, high efficiency and ideal repeatability and economical efficiency, etc.; moreover, the extracted DNA has high concentration, purity and yield, and can be directly used in subsequent molecular biology experimental researches such as PCR amplification without purification.

The invention discloses a qPCR (quantitative Polymerase Chain Reaction) quantitative standard for detecting library DNA (Deoxyribonucleic Acid) concentration of aneuploids, i.e., 21 trisome, 18 trisome and 13 trisome of fetal chromosomes based on a semiconductor sequencing method and a preparation method therefor and belongs to the field of noninvasive prenatal detection on fetal aneuploids. The standard is used for carrying out quality control on detection concentration of detected samples. The preparation method for the qPCR quantitative standard disclosed by the invention comprises five steps: (1) synthesizing joint sequences and primers; (2) preparing simulated fetal free DNA libraries; (3) amplifying and purifying the libraries; (4) producing a library standard curve; and (5) carrying out sample detection. According to the method disclosed by the invention, the DNA concentration of sample libraries can be rapidly determined, and a guarantee of accuracy and validity of noninvasive prenatal detection on the aneuploids, i.e., the 21 trisome, the 18 trisome and the 13 trisome of the fetal chromosomes is provided.

The invention discloses a kit and a method for quickly extracting DNA (deoxyribonucleic acid) from colla corii asini. A combined technology of an SDS (sodium dodecyl sulfate)-protease K and Chelex-100 combined process DNA extraction technology and a Glassmilk DNA purification technology is applied to DNA extraction of the colla corii asini. The dosage of needed colla corii asini paste is controlled to be 0.3ml to 0.5ml, the weight of colla corii asini blocks can be controlled to be 0.2mg to 0.6mg, and extracted DNA is high in concentration, excellent in purity, stable and difficult to degrade; due to extracted DNA, sequence fragments of 100bp-500bp can be amplified, and subsequent molecular biology experiments of gene cloning, sequencing and the like can be carried out. According to the method, the operation is simple, the dosage is low, and the consumed time is short; the method is especially suitable for DNA extraction of various types of colla corii asini products with low nucleic acid content and high degradation degree caused by deep processing; a foundation is provided for realizing colla corii asini source identification by adopting a DNA molecular biology identification technology.

The invention discloses an aneuploidy biological information analysis method and system, wherein the method comprises: 1) constructing a reference database; 2) calculating UR ratio; 3) constructing statistical parameters of the reference database; 4) calculating Z value; 5) decreasing fetal chromosomal aneuploidy false positives due to microdeletion or micro-duplication present in maternal chromosomes according to the above process of decreasing fetal chromosomal aneuploidy false positives due to microdeletion or micro-duplication present in maternal chromosomes; 6) constructing a fetal DNA concentration prediction model to predict fetal DNA concentration according to the above process of constructing a fetal DNA concentration prediction model; 7) calculating percentage of DNA of each fetal chromosome to total DNA; 8) judging autosomal aneuploidy; 9) judging allosomal anomalies. By using the method and system according to the technical scheme, analytical accuracy is improved greatly.

The invention provides a simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples. According to the simple method, the DNA in environmental samples is fully released and exposed by using combination of methods such as glass bead grinding, liquid nitrogen repetitive freeze-thawing, SDS (sodium-dodecyl sulphate), lysozyme and a protease-K method for the different soil samples, the DNA purification is carried out through multiple phenol imitation extraction processes, and the determination of the DNA extraction efficiency is carried out by adding an internal standard bacterial strain Xcc8004. The simple method has simple requirements on sample pretreatment; the quantity of the required environmental samples is small; the obtained DNA has relatively high concentration and purity; the extraction efficiency is relatively high; the analysis and detection of antibiotic resistance gene pollution in the complex environmental samples can be met; and the method is simple and reliable and has practical application value.

The invention relates to a biological analysis method, especially to an analysis method for detecting the diversity of soil microorganism. The method comprises the following steps of: (1) carrying out DNA extraction in soil; (2) detecting DNA concentration and purity in soil by using the DNA extract obtained from Step (1); (3) carrying out PCR specific amplification on the extracted DNA from soil; and (4) carrying out denaturing gradient gel electrophoresis analysis on the PCR reaction product. By the adoption of the method for detection and analysis of various soils, the result is consistent with the real state of microorganisms as there is no loss of the extracted DNA solution during the treatment process. Therefore, the invention provides a perfect solution for the research of the soil microorganism state.

The invention relates to a method for electrochemically detecting concentration of specific single-stranded DNA based on exonuclease and a nucleic acid probe and belongs to the technical field of analytical chemistry. The invention designs and synthesizes two types of hairpin-type probes P1 and P2. The method comprises the following steps of: firstly modifying a gold electrode with gold nanoparticles by adopting 1,6-hexanedithiol (HDT), preparing an AuNPs-HDT-Au electrode, and then modifying the probe P1 onto the electrode; taking a specific single stranded target DNA as a to-be-detected object, so that the probes P1 and P2 can coexist when no target DNA exists, and the target DNA can trigger two independent reaction cycles when the target DNA, the probe P2 and the exonuclease ExoIII exist; and finally when heme exists, generating a strong signal under the interaction of a G-tetraplex sequence of the probe P1 on the surface of the electrode and heme, and detecting the target DNA by adopting a differential pulse voltammetry, wherein a peak current signal and the target DNA concentration are correlated in a certain concentration range, so that detection on the target DNA concentration is realized. The method provided by the invention has the advantages of high sensitivity and strong specificity.

The invention discloses a molecular detecting method of pine grease ulcer germ, which is characterized by the following: utilizing primer designing software Prime 5.1 to design a couple of general prime G1/G2 and astopic prime S1/S2 on the fusarium IGS segment; adopting pine germ sample DNA extract as form; proceeding the first-turn PCR augmenting reaction of general prime; diluting the PCR augmented product by 100 times; fetching 1 mul as form to proceed the second-turn astopic prime PCR augmenting reaction; proceeding electrophoretic detection for augmented product; affirming the detected germ as pine grease ulcer germ if the DNA astopic band with 873bp exists. The invention needs only one working day to finish detect, whose density of reaches minimum quantity germ DNA is 5*10-3pg/ mul and conidiospore is 10units/100 mul.

The invention discloses a method for acquiring the concentration of fetal cell-free DNA. For the length data of cfDNA fragments in maternal peripheral blood, a Gaussian mixture model with K normal distributions is adopted to quantize maternal and fetal cfDNA, five wave crests and corresponding distribution ranges are automatically and accurately acquired, and then the concentration of fetal cfDNA is obtained. The method provides more suitable and more reliable concentration of fetal cfDNA for non-invasive prenatal testing (NIPT) and has universality and accuracy. The method is effective to both male fetuses and female fetuses as the distribution areas of lengths of maternal and fetal cfDNA fragments are dynamically determined; besides, the distribution of maternal and fetal cfDNA can be automatically acquired and recognized for samples of different fetuses of different gestational ages, and thus the accuracy of the concentration of fetal cfDNA is guaranteed.

The invention discloses a simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method universally applied to chrysanthemum and related species thereof. In a PCR reaction system, the primer concentration is 0.32 MuM/Mul; the Taq enzyme concentration is 0.2 U/Mul; the dNTP concentration is 0.4 MuM/Mul; the Mg<2+> concentration is 0.65 MuM/Mul; and the DNA concentration is 2 to 4 ng/Mul. Stripes obtained by the method for performing SSR molecular marker PCR reaction are clear and have high polymorphism and high repeatability. The method is simple in operation and commonly used in wild species and varieties of chrysanthemum, Ajania, Opisthopappus Shih and Crossostephium Less, and has a wide application prospect.

The invention provides a feces preservation solution, a preparation method thereof and a feces preservation method. The feces preservation solution contains 0.10-0.20 M of Tris base, 0.25-0.75 M of ethylenediaminetetraacetic acid (EDTA), 10 mM-room temperature saturated sodium chloride (NaCl), 5-15% of dimethyl sulfoxide (DMSO), 2.5-10% of ethanol and 5-20% of glycerol, wherein the pH of the fecespreservation solution is 7.0-9.0. The feces preservation solution can stably preserve microbial diversity in feces samples at normal temperature and improve DNA concentration, can ensure the accuracyof sequencing analysis of intestinal flora, realizes normal-temperature preservation and transportation of the feces samples, has a good preservation effect compared with an existing preservation solution, does not contain toxic components such as guanidine thiocyanate, and is safer to operate.

The invention discloses a method for detecting rearrangement clonality of correlative genes of lymphocyte and belongs to the technical field of biology. According to the method, a high-throughput next-generation sequencing method is adopted for detecting rearrangement clonality of the correlative genes of lymphocyte tissue, and specifically, a multifunctional primer and a PCR reagent are utilizedto conduct multiplication on samples to be detected at first to obtain a multi-sample gene locus multiplication sublibrary, wherein both ends of the multiplication sublibrary are connected with DNA segments of different connectors, one side of the multiplication sublibrary is a sequencing connector which can contain a sample tag for distinguishing sequencing results of the different samples, and the other side of the multiplication sublibrary is a fixed connector used for connecting caught particles; then, high-throughput sequencing is conducted on the multiplication sublibrary, sequencing results are subjected to classified sorting according to a tag sequence, the samples and target genetic loci, and whether or not there is clonality rearrangement of target genes is analyzed. The method has the advantages that the detection throughput, the sensitivity and the specificity are high, the sensitivity can reach 0.01%, and the DNA concentration of the samples can be lowered to 0.1-1ng/microliter.

The invention relates to a method for detecting DNA (deoxyribonucleic acid) through high sensitivity Raman spectrum. According to the method, chains on the surfaces of magnetic beads are used for substituting the potentiation of polymerization and silver so as to realize dual amplification of the signal. The magnetic beads are used for fixing molecular beacons and separating chain substituted products. After hybridization of the target DNA and the molecular beacons, beacon rings are opened, raman dye-silver nano-composite modified by a primer is connected with the magnetic beads through hybridization of the primer and the stem end of the beacon so as to induce the DNA chain to carry out polymerization reaction to release target molecules, and the released DNA molecules and the molecular beacon of the other magnetic bead are hybridized so as to generate the other polymerization circulation. After the circulation chain substitutes polymerization, a separation chain substitutes the product, a silver nanoparticle shell is deposited on the surface of the nano-composite, so as to enhance the raman signal of the dye. The correlation between the raman signal and the concentration and sequence of the target DNA are used for DNA analysis. The concentration range of DNA detected by the method is 10-13-10-8mol L-1, and the method can be used for distinguishing completely matched target DNA and single-base mismatch DNA, and has certain application value.

The invention belongs to the field of biotechnology and particularly provides a Periplaneta Americana L. specific COI primer, a kit containing the same and application of the Periplaneta Americana L. specific COI primer. According to a conserved sequence of Periplaneta Americana L. mtDNA, the invention designs a pair of the Periplaneta Americana L. specific COI primers JmF1 and JmR1 (such as SEQ ID No. 1 and SEQ ID No. 2), and detected that, the primer has the specific amplification ability to Periplaneta Americana L. only, the size of an amplified product is 452bp, and a sensitivity DNA concentration detection limit is 0.390625ng/uL. According to the Periplaneta Americana L. specific COI primer, the kit and the application, by utilizing a PCR detection technology, a specific single band is obtained through specific primer amplification, operation is simple, and the Periplaneta Americana L. specific COI primer has the characteristics of high efficiency, low cost and strong specificity and improves the accuracy of detection.

The invention discloses a device for detecting the aneuploidy of fetus chromosomes based on single-sample peripheral blood. The device comprises a sequencing data processing unit and a result analysis unit. Whether the chromosomes to be detected are aneuploid chromosomes or not are judged by comparing the quantity of short sequences of 101-150 bp with the quantity of long sequences of 151-200 bp on the same chromosome. According to the device, self control of samples is conducted, and accuracy is high; the dependency of fetus free DNA concentration is small, and the false-negative problem of the aneuploidy of the chromosomes under the condition of low fetus free DNA concentration is solved to a great extent; moreover, the requirement for the sequencing data size is low compared with an original method, and thus cost can be further lowered.

The invention belongs to the field of biology, and discloses a body fluid suspension cell DNA extracting kit. The extracting kit consists of a resuspension solution, a protease K solution, an RNA enzyme A solution, lysate, a precipitant, a washing solution A, a washing solution B and a DNA dissolved solution. A method for extracting body fluid suspension cell DNA by virtue of the kit comprises the following steps: conducting centrifuging, precipitating and re-suspending, adding the lysate, promoting lysis of the protease K solution, adding the precipitant and conducting centrifuging, adding the washing solution A, conducting centrifuging and washing with the washing solution B, and conducting drying at room temperature and adding the DNA dissolved solution. Different from an existing DNA extracting method or DNA extracting kit which is expensive, simple in object of using and relatively poor in integrity of extracted DNA and which are not applicable to extraction of the body fluid suspension cell DNA, the method and the kit provided by the invention are cheap, relatively high in concentration of the extracted DNA and good in integrity of the extracted DNA and are applicable to extraction of various body fluid suspension cell DNA.