CN112501282A - Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection - Google Patents
Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection Download PDFInfo
- Publication number
- CN112501282A CN112501282A CN202011596447.1A CN202011596447A CN112501282A CN 112501282 A CN112501282 A CN 112501282A CN 202011596447 A CN202011596447 A CN 202011596447A CN 112501282 A CN112501282 A CN 112501282A
- Authority
- CN
- China
- Prior art keywords
- dna
- whole blood
- adsorption column
- bsmi
- base sequences
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and carrying out qPCR amplification detection, 1) sequencing data processing, 2) Seurat data filtering, 3) cell clustering and 4) cell population marker gene analysis.
Description
Technical Field
The invention relates to the technical field of gene detection, in particular to a method for obtaining base sequences of BsmI and PvuII sites by extracting DNA from whole blood and performing qPCR amplification detection.
Background
Osteoporosis is a metabolic disease of the whole body bone, and is mainly characterized by a decrease in bone mass and a change in bone microstructure accompanied by a decrease in bone strength, an increase in bone fragility, an increase in fracture risk, and the like. According to epidemiological statistics, the prevalence rate of osteoporosis in people over 50 years old in China is 20.7% for women and 14.4% for men. And according to global epidemiological investigation, the prevalence of osteoporosis is high in all countries. The onset of osteoporosis is associated with a variety of factors, of which sex factors and age factors are the most relevant to the onset of osteoporosis. The pain and the fracture caused by the osteoporosis seriously affect the life quality of the patient and endanger the survival of the patient, and the existing treatment means has relatively limited effect on the osteoporosis. In the elderly women, after the pathological fracture is mostly followed by osteoporosis, the residual fatality rate caused by osteoporosis fracture is high, and the fatality rate of the osteoporosis fracture exceeds the sum of the fatality rates of the elderly women suffering from breast cancer, cervical cancer and uterine body cancer at present.
The current clinical methods for diagnosing osteoporosis mainly comprise: 1. the physician consults a past medical history which provides a high risk factor for osteoporosis, such as alcohol abuse, heavy smoking, pre-menopausal bilateral ovariectomy, etc., and then needs further examination. 2. Bone mass is measured based on the measurement of bone mineral content and bone density, but the occurrence of fractures and osteoporosis does not depend solely on bone mass. 3. Ultrasound bone determination measures the speed of ultrasound through bone tissue, amplitude attenuation and stiffness index reflecting bone structure and bone mass.
The existing methods for diagnosing osteoporosis mainly have the following defects: 1. the physician can only provide reference for inquiring the past medical history and can not accurately judge whether osteoporosis exists. 2. Bone mass measurement is a commonly used means at present, but whether fracture occurs or not is mainly determined by bone strength, and the bone strength is composed of various aspects such as bone mass, bone quality, bone composition and bone shape, so that the judgment by the bone mass alone is not accurate. 3. The result of ultrasonic measurement is not the bone density or the bone mineral content, and can not compare with the true value, needs more observation data, and the huge inconvenience of moving of ultrasonic equipment in addition is higher shortcoming such as the cost of each measurement. Therefore, an improved product for diagnosing osteoporosis is needed in the market.
Disclosure of Invention
The invention provides a method for obtaining base sequences of BsmI and PvuII sites by extracting DNA from whole blood and performing qPCR amplification detection.
The scheme of the invention is as follows:
a method for extracting DNA from whole blood to carry out qPCR amplification detection to obtain base sequences of BsmI and PvuII sites comprises the following steps:
1) preparing a required DNA sample, extracting the DNA of the sample from whole blood, and storing the obtained DNA of the sample at the temperature of between 20 ℃ below zero and 4 ℃ for later use;
2) qPCR amplification of sample DNA, namely diluting the sample DNA in the step 1) to 5-30 ng/mu L, constructing a reaction system in a PCR tube, sending the reaction system into a PCR amplification instrument, setting a reaction program for reaction, and obtaining a target gene after the reaction is finished;
3) and (3) detecting the locus, namely detecting the target gene in the step 2) to obtain a detection report.
And (3) judging the risk of osteoporosis by detecting the base sequences of the BsmI site and the PvuII site.
As a preferred technical scheme, the DNA of the whole blood extracted sample is as follows:
cutting a Tip of a 1-mL Tip, inserting a pipette, extending into a blood collection tube, repeatedly blowing and beating whole blood for 3-4 times, fully suspending precipitated white blood cells, transferring the whole blood to an EP tube, adding MGS lysate, adding proteinase K, uniformly mixing, heating in a water bath kettle at 60 ℃ for 10-15 min, wherein the uniformly mixing time is more than 2 times;
taking out the EP pipe from the water bath kettle, pouring all solution in the EP pipe into a centrifugal adsorption column, putting the adsorption column into a collecting pipe, centrifuging at 8000rpm for 1min, taking out the adsorption column, and pouring out waste liquid in the collecting pipe;
adding cleaning solution into the adsorption column
8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring off the waste liquid in the collection tube;
rinsing twice, taking out the adsorption column after the completion, pouring off the waste liquid in the collection tube, putting the adsorption column back into the collection tube again, centrifuging at 12000rpm for 1min, and removing the residual ethanol;
placing the adsorption column in a new 1.5ml centrifuge tube, and drying at room temperature for 2-3 min; dripping 60 deg.C eluent ET into the adsorption column near the center of the adsorption membrane, centrifuging at 12000rpm for 1-2min, collecting the liquid in the tube as sample DNA solution, covering with a cover, and lightly pulling the liquid in the tube to mix well.
As a preferable technical scheme, the rinsing treatment is to add a rinsing liquid Wash into the adsorption column at 8000rpm, centrifuge for 30s, take out the adsorption column and pour the waste liquid in the collection tube.
As a preferable technical scheme, the reaction system in the step 2) is as follows:
as a preferred technical scheme, MGB-TaqMan-FAM adopts FAM6 at the Apa I site, and the rest is FAM.
As a preferable technical scheme, the reaction procedure in the step 2) is as follows:
S3-S4 were cycled 45 times.
The method for obtaining the base sequences of BsmI and PvuII sites by extracting DNA from whole blood and performing qPCR amplification detection is adopted in the technical scheme, 1) the required DNA sample is prepared, the sample DNA is extracted from the whole blood, and the obtained sample DNA is stored at the temperature of-20-4 ℃ for later use; 2) qPCR amplification of sample DNA, namely diluting the sample DNA in the step 1) to 5-30 ng/mu L, constructing a reaction system in a PCR tube, sending the reaction system into a PCR amplification instrument, setting a reaction program for reaction, and obtaining a target gene after the reaction is finished; 3) and (3) detecting the locus, namely detecting the target gene in the step 2) to obtain a detection report.
The invention has the advantages that: the method has the advantages that the operation steps are simple, the graph result is visual, the interpretation is easy, the detection time is shortened, the detection efficiency is improved, and the detection accuracy is improved;
provides effective information, thereby improving the prediction accuracy of the osteoporosis risk.
Drawings
FIG. 1 is a graph of the amplification of the VDR BsmI detection site GG in accordance with the present invention;
FIG. 2 is a graph showing the AG amplification of VDR BsmI detection sites in accordance with one embodiment of the present invention;
FIG. 3 is a graph showing the AA amplification of the VDR BsmI detection site in the embodiment of the present invention;
FIG. 4 is a graph showing the amplification of the ESR PvuII detection site AA in the example of the present invention;
FIG. 5 is a graph showing the amplification of AG at the detection site of ESR PvuII in the example of the present invention;
FIG. 6 is a graph showing the amplification of the ESR PvuII detection site GG in the example of the present invention.
FIG. 7 is an analytical flow chart according to the present invention;
FIG. 8 is an interface diagram of the Roche 480 analysis software analysis site selection analysis according to an embodiment of the present invention;
FIG. 9 is an interface diagram of a Roche 480 analysis software selection analysis sample according to an embodiment of the present invention;
FIG. 10 is a graphical illustration of a Roche 480 analysis software analysis parameters selection interface according to an embodiment of the present invention;
FIG. 11 is a FAM interface diagram of the Roche 480 analysis software selection channel according to an embodiment of the present invention;
FIG. 12 is an interface diagram of FAM analysis results of Roche 480 analysis software according to an embodiment of the present invention;
FIG. 13 is an interface diagram of a Roche 480 analysis software exchange analysis channel according to an embodiment of the present invention;
FIG. 14 is a VIC channel interface diagram of Roche 480 analysis software according to an embodiment of the present invention;
FIG. 15 is an interface diagram of Roche 480 analysis software for importing analysis channels into the reading software separately according to an embodiment of the present invention;
FIG. 16 is an interface diagram of Roche 480 analysis software showing the amplification map of the FAM channel in accordance with an embodiment of the present invention;
FIG. 17 is an interface diagram of the Roche 480 analysis software showing the amplification map of the VIC channel in accordance with one embodiment of the present invention;
FIG. 18 is an interface diagram of the Roche 480 analysis software showing FAM and VIC dual channel amplification maps in accordance with the present invention.
Detailed Description
In order to make up for the above deficiencies, the present invention provides a method for obtaining base sequences of BsmI and PvuII sites by extracting DNA from whole blood and performing qPCR amplification detection to solve the above problems in the background art.
A method for extracting DNA from whole blood to carry out qPCR amplification detection to obtain base sequences of BsmI and PvuII sites comprises the following steps:
1) preparing a required DNA sample, extracting the DNA of the sample from whole blood, and storing the obtained DNA of the sample at the temperature of between 20 ℃ below zero and 4 ℃ for later use;
2) qPCR amplification of sample DNA, namely diluting the sample DNA in the step 1) to 5-30 ng/mu L, constructing a reaction system in a PCR tube, sending the reaction system into a PCR amplification instrument, setting a reaction program for reaction, and obtaining a target gene after the reaction is finished;
3) and (3) detecting the locus, namely detecting the target gene in the step 2) to obtain a detection report.
And (3) judging the risk of osteoporosis by detecting the base sequences of the BsmI site and the PvuII site.
The whole blood extracted sample DNA is as follows:
cutting a Tip of a 1-mL Tip, inserting a pipette, extending into a blood collection tube, repeatedly blowing and beating whole blood for 3-4 times, fully suspending precipitated white blood cells, transferring the whole blood to an EP tube, adding MGS lysate, adding proteinase K, uniformly mixing, heating in a water bath kettle at 60 ℃ for 10-15 min, wherein the uniformly mixing time is more than 2 times;
taking out the EP pipe from the water bath kettle, pouring all solution in the EP pipe into a centrifugal adsorption column, putting the adsorption column into a collecting pipe, centrifuging at 8000rpm for 1min, taking out the adsorption column, and pouring out waste liquid in the collecting pipe;
adding cleaning solution into the adsorption column
8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring off the waste liquid in the collection tube;
rinsing twice, taking out the adsorption column after the completion, pouring off the waste liquid in the collection tube, putting the adsorption column back into the collection tube again, centrifuging at 12000rpm for 1min, and removing the residual ethanol;
placing the adsorption column in a new 1.5ml centrifuge tube, and drying at room temperature for 2-3 min; dripping 60 deg.C eluent ET into the adsorption column near the center of the adsorption membrane, centrifuging at 12000rpm for 1-2min, collecting the liquid in the tube as sample DNA solution, covering with a cover, and lightly pulling the liquid in the tube to mix well.
And the rinsing treatment comprises the steps of adding a rinsing liquid Wash into the adsorption column at 8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring out waste liquid in the collecting pipe.
The reaction system in the step 2) is as follows:
the MGB-TaqMan-FAM adopts FAM6 at the Apa I site, and the balance is FAM.
As a preferable technical scheme, the reaction procedure in the step 2) is as follows:
S3-S4 were cycled 45 times.
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments.
Example (b):
extraction of DNA from whole blood
1. Cutting off a Tip of a 1-mL Tip head, inserting a pipette, extending into a blood collection tube, repeatedly blowing and beating whole blood for 3-4 times to fully suspend precipitated white blood cells, transferring 200 mul of whole blood into a 2mL EP tube, adding 200 mul of MGS lysate and 20 mul of proteinase K, uniformly mixing, heating in a 60 ℃ water bath kettle for 10min (no more than 15min), uniformly mixing at least for 2-3 times, otherwise, enabling hemoglobin to cohere and agglomerate.
2. Taking out the sample from the water bath, pouring the solution into a centrifugal adsorption column, placing the adsorption column into a collecting tube, centrifuging at 8000rpm for 1min, taking out the adsorption column, and pouring off the waste liquid in the collecting tube.
3. Adding 500 μ l of cleaning solution into the adsorption column
8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring the waste liquid in the collecting pipe.
4. Adding 500 μ l of rinsing liquid Wash to the adsorption column at 8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring off waste liquid in the collection tube.
5. Repeat 4 times.
6. Taking out the adsorption column, pouring off waste liquid in the collection tube, putting the adsorption column back into the collection tube again, and centrifuging at 12000rpm for 1min to remove residual ethanol as much as possible.
7. The adsorption column was placed in a new 1.5ml centrifuge tube and dried for 2-3min at room temperature.
8. And (3) suspending and dropwise adding 50-100 μ l of eluent ET (preheating the eluent ET at 65 ℃ in advance to improve the elution efficiency) into the adsorption column close to the center of the adsorption film. 12000rpm, centrifuging for 1-2min, and collecting liquid in the centrifugal tube to obtain the DNA solution. And lightly stirring the liquid in the centrifugal tube after covering the cover to uniformly mix. The sample DNA is stored at 4 ℃ or-20 ℃ for a long period of time according to the application.
9. Can be directly used for PCR, enzyme digestion, library construction, sequencing reaction and the like.
qPCR amplification
Diluting the qualified DNA sample to 5-30 ng/muL;
qPCR reaction system
| Reagent | Volume of |
| 2 × qPCR Mix (New sea NuHi SNP Mix) | 5μL |
| Primer F/R | 0.4μL |
| MGB-TaqMan-FAM ( |
0.2μL |
| MGB-TaqMan-VIC | 0.2μL |
| ddH2O | 3.2μl |
| Template DNA | 1μL |
Interpreting the result according to the result interpretation standard and issuing a detection report; the Roche 480 analysis software operates as in FIGS. 8-18 to analyze the amplification curves;
the risk of osteoporosis is judged by detecting the base sequences of BsmI site and PvuII site, which are shown in figure 1 to figure 6;
osteoporosis risk interpretation
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (6)
1. A method for extracting DNA from whole blood to carry out qPCR amplification detection to obtain base sequences of BsmI and PvuII sites is characterized by comprising the following steps:
1) preparing a required DNA sample, extracting the DNA of the sample from whole blood, and storing the obtained DNA of the sample at the temperature of between 20 ℃ below zero and 4 ℃ for later use;
2) qPCR amplification of sample DNA, namely diluting the sample DNA in the step 1) to 5-30 ng/mu L, constructing a reaction system in a PCR tube, sending the reaction system into a PCR amplification instrument, setting a reaction program for reaction, and obtaining a target gene after the reaction is finished;
3) and (3) detecting the locus, namely detecting the target gene in the step 2) to obtain a detection report.
2. The method for obtaining the base sequences of BsmI and PvuII sites by qPCR amplification assay of whole blood extracted DNA as claimed in claim 1, wherein the DNA of the whole blood extracted sample is as follows:
cutting a Tip of a 1-mL Tip, inserting a pipette, extending into a blood collection tube, repeatedly blowing and beating whole blood for 3-4 times, fully suspending precipitated white blood cells, transferring the whole blood to an EP tube, adding MGS lysate, adding proteinase K, uniformly mixing, heating in a water bath kettle at 60 ℃ for 10-15 min, wherein the uniformly mixing time is more than 2 times;
taking out the EP pipe from the water bath kettle, pouring all solution in the EP pipe into a centrifugal adsorption column, putting the adsorption column into a collecting pipe, centrifuging at 8000rpm for 1min, taking out the adsorption column, and pouring out waste liquid in the collecting pipe;
adding cleaning solution into the adsorption column
8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring off the waste liquid in the collection tube;
rinsing twice, taking out the adsorption column after the completion, pouring off the waste liquid in the collection tube, putting the adsorption column back into the collection tube again, centrifuging at 12000rpm for 1min, and removing the residual ethanol;
placing the adsorption column in a new 1.5ml centrifuge tube, and drying at room temperature for 2-3 min; dripping 60 deg.C eluent ET into the adsorption column near the center of the adsorption membrane, centrifuging at 12000rpm for 1-2min, collecting the liquid in the tube as sample DNA solution, covering with a cover, and lightly pulling the liquid in the tube to mix well.
3. The method of claim 1 for obtaining base sequences of BsmI and PvuII sites by qPCR amplification detection of DNA extracted from whole blood, wherein: and the rinsing treatment comprises the steps of adding a rinsing liquid Wash into the adsorption column at 8000rpm, centrifuging for 30s, taking out the adsorption column, and pouring out waste liquid in the collecting pipe.
4. The method for obtaining base sequences of BsmI and PvuII sites by qPCR amplification detection of DNA extracted from whole blood as claimed in claim 1, wherein the reaction system in step 2) is as follows:
5. the method of claim 4 for obtaining base sequences of BsmI and PvuII sites by qPCR amplification detection of DNA extracted from whole blood, wherein: the MGB-TaqMan-FAM adopts FAM6 at the Apa I site, and the balance is FAM.
6. The method for obtaining base sequences of BsmI and PvuII sites by qPCR amplification assay of DNA extracted from whole blood as claimed in claim 5, wherein the reaction procedure in step 2) is as follows:
S3-S4 were cycled 45 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011596447.1A CN112501282A (en) | 2020-12-29 | 2020-12-29 | Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011596447.1A CN112501282A (en) | 2020-12-29 | 2020-12-29 | Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112501282A true CN112501282A (en) | 2021-03-16 |
Family
ID=74951866
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011596447.1A Pending CN112501282A (en) | 2020-12-29 | 2020-12-29 | Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112501282A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220426A (en) * | 2018-02-27 | 2018-06-29 | 北京岳昊科技发展有限公司 | A kind of kit for being used to detect osteoporosis |
| CN110257507A (en) * | 2019-07-11 | 2019-09-20 | 上海联吉医学检验所有限公司 | A kind of pregnancy period folic acid metabolism, calcium metabolism and H-type hypertension associated detecting method |
| CN111019940A (en) * | 2019-12-25 | 2020-04-17 | 上海派森诺生物科技股份有限公司 | Extracting solution for directly extracting whole blood genome DNA and extracting method thereof |
| CN111534588A (en) * | 2020-04-23 | 2020-08-14 | 南京实践医学检验有限公司 | Kit and method for detecting gene mutation in acute lymphocytic leukemia based on fluorescent quantitative PCR |
| CN111996244A (en) * | 2020-10-28 | 2020-11-27 | 上海鼎晶生物医药科技股份有限公司 | Composition for detecting single nucleotide polymorphism and application thereof |
-
2020
- 2020-12-29 CN CN202011596447.1A patent/CN112501282A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220426A (en) * | 2018-02-27 | 2018-06-29 | 北京岳昊科技发展有限公司 | A kind of kit for being used to detect osteoporosis |
| CN110257507A (en) * | 2019-07-11 | 2019-09-20 | 上海联吉医学检验所有限公司 | A kind of pregnancy period folic acid metabolism, calcium metabolism and H-type hypertension associated detecting method |
| CN111019940A (en) * | 2019-12-25 | 2020-04-17 | 上海派森诺生物科技股份有限公司 | Extracting solution for directly extracting whole blood genome DNA and extracting method thereof |
| CN111534588A (en) * | 2020-04-23 | 2020-08-14 | 南京实践医学检验有限公司 | Kit and method for detecting gene mutation in acute lymphocytic leukemia based on fluorescent quantitative PCR |
| CN111996244A (en) * | 2020-10-28 | 2020-11-27 | 上海鼎晶生物医药科技股份有限公司 | Composition for detecting single nucleotide polymorphism and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105349654B (en) | A kind of probe for detecting EGFR genetic mutation, primer, detection architecture and kit | |
| CN107663533A (en) | A kind of lung cancer EGFR L858R and 19Del ddPCR detection methods and application | |
| CN104004840B (en) | Test kit for early screening Yu diagnosis of prostate cancer | |
| WO2020048518A1 (en) | Group of genes for molecular typing of medulloblastoma and use thereof | |
| CN109457032B (en) | Thyroid cancer molecular diagnosis kit | |
| CN108823305B (en) | Multiplex PCR method for synchronously detecting 8 tumor-associated long-chain non-coding RNAs | |
| CN110423816A (en) | Prognosis in Breast Cancer quantitative evaluation system and application | |
| CN107435062B (en) | Peripheral blood gene marker for discriminating benign and malignant pulmonary nodules and application thereof | |
| CN115851951A (en) | Construction of early liver cancer detection model containing multiple groups of chemical marker compositions and kit | |
| CN112501282A (en) | Method for obtaining BsmI and PvuII site base sequences by extracting DNA from whole blood and performing qPCR amplification detection | |
| CN116121384A (en) | Gene methylation marker combination and screening model for screening cervical high-level lesions | |
| CN110317870B (en) | miRNA marker for diagnosing prostate cancer, kit, system, application and detection method | |
| CN112609002B (en) | Peripheral blood miRNA colon cancer diagnosis marker combination and detection kit thereof | |
| CN108753981A (en) | Application of the quantitative detection of HOXB8 genes in colorectal cancer Index for diagnosis | |
| CN112695095A (en) | Peripheral blood miRNA lung cancer diagnosis marker combination and detection kit thereof | |
| CN114381519A (en) | Serum miRNA marker combination for breast cancer auxiliary diagnosis and detection kit thereof | |
| CN107858425A (en) | Applications and detection method of the miRNA 4741 as primary hepatic carcinoma diagnosis mark | |
| CN113470754A (en) | Gene marker for tumor prognosis evaluation, evaluation product and application | |
| RU2522923C1 (en) | Integrated method for detecting circulating tumour cells in blood of patients suffering from breast cancer | |
| CN111690746A (en) | Platelet RNA marker related to lung cancer and application thereof | |
| WO2020135422A1 (en) | Health risk assessment method | |
| CN112011610A (en) | Kit for cancer prognosis detection | |
| CN111826445B (en) | Application of miR-1468-5p in evaluation of expression of PD-L1 of cervical cancer patient | |
| CN116516008B (en) | Gastric mucosa intestinal epithelium metaplasia marker JUN and application thereof | |
| CN117265112B (en) | Digital micro-droplet RNA amplification detection system for prostate cancer and application of PCA3 gene in system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |