CN111378746A - Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene - Google Patents
Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene Download PDFInfo
- Publication number
- CN111378746A CN111378746A CN201811598534.3A CN201811598534A CN111378746A CN 111378746 A CN111378746 A CN 111378746A CN 201811598534 A CN201811598534 A CN 201811598534A CN 111378746 A CN111378746 A CN 111378746A
- Authority
- CN
- China
- Prior art keywords
- molecular beacon
- sequence
- kit
- casp7
- detecting
- 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
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- 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
Abstract
The invention discloses a typing detection method and a kit for rs12415607 site polymorphism of CASP7 gene. The invention adopts primers SEQ1 and SEQ2 with CASP7 gene specificity to amplify a CASP7 gene fragment, and designs CASP7 gene specificity artificial simulation nucleic acid molecular beacons SEQ3-FAM and SEQ4-VIC in an amplification region defined by the primers with CASP7 gene specificity. The method for judging the rs12415607 site polymorphism of the CASP7 gene based on the gene specificity PCR combined with the artificial simulated nucleic acid molecular beacon, provided by the invention, has the advantages of high accuracy, high detection speed, simplicity in operation, objective result interpretation, less closed-tube reaction pollution and the like, and is very suitable for large-scale clinical development.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a typing detection method and a kit for rs12415607 site polymorphism of CASP7 gene.
Background
Lung cancer is one of the leading death factors associated with cancer worldwide, and although smoking is a significant cause of lung cancer, only 20% of smokers will eventually develop lung cancer in practice. This suggests that genetic factors also play a crucial role in the pathogenesis of lung cancer. In the etiology research of lung cancer, a few susceptibility genes and low-penetrance genes related to the lung cancer have been identified. In addition, other pathways are likely to be involved in lung cancer pathogenesis, where the role of apoptotic genes in human cancer susceptibility is becoming more and more important.
Apoptosis plays an important role in development and cell and tissue homeostasis. Defects in apoptotic pathways may cause accumulation of somatic mutations, thereby increasing the risk of cancer. Cell destruction during apoptosis is achieved by Caspases (CASPs), a family of highly conserved caspases. Based on their pro-apoptotic function, caspases can be isolated as the promoter CASP, which links death signals to cell death programs; CASP is performed in a coordinated proteolytic process leading to the destruction of critical cellular structures. In humans, there are two executions CASP, CASP3 and CASP 7. The risk of lung cancer of an individual with AA genotype at the single nucleotide polymorphism locus rs12415607 (A/C) in CASP7 is obviously increased. The risk of lung cancer can be analyzed on the whole by detecting the locus, thereby providing a reference for preventing the disease.
At present, methods for detecting gene polymorphism mainly include a PCR-Sanger sequencing method, a chip hybridization method, a high-resolution melting curve method and the like. Although these methods can detect gene polymorphisms to some extent, they have considerable limitations. The Sanger sequencing method has more steps, needs PCR post-treatment, is complex to operate, is easy to cause pollution, and cannot meet clinical requirements. The chip hybridization method is complicated in operation, and detection thereof depends on expensive equipment and instruments, resulting in high cost. The high-resolution dissolution curve method has high requirements on instruments, can be used only by a machine which is provided with high-resolution software and is sensitive to temperature, and has difficulty in clinical popularization. The fluorescent quantitative PCR based on the Taqman hydrolysis probe cuts off the probe to generate a fluorescent signal by utilizing the exonuclease activity of Taq enzyme, and the fluorescent quenching is not thorough due to the fact that a fluorescent group and a quenching group of the Taqman probe are not close to each other closely, and a background fluorescent signal exists. In addition, the Taqman probe has poor single base mismatch recognition capability, easily generates a non-specific fluorescent signal, interferes result interpretation, and further influences the detection accuracy. Therefore, a simple, convenient, high-sensitivity, accurate and reliable method for detecting gene polymorphism is urgently needed clinically.
The Molecular Beacon (Molecular Beacon) is in a hairpin type in spatial structure and consists of a circular region and a stem region, wherein the circular region is complementary with a target DNA sequence and is about 15-35 nucleotides long, the stem region is about 5-7 nucleotides long, the stem region is formed by a complementary sequence which has higher GC content and is irrelevant with the target sequence, and the 5 'end of the Molecular Beacon is marked with a fluorescent group (F) and the 3' end of the Molecular Beacon is marked with a quenching group (Q). In the case of molecular beacons, the fluorescent group is close to the quencher group (about 7-10 nm) in the free state. At the moment, fluorescence resonance energy transfer occurs, so that fluorescence emitted by the fluorescent group is absorbed by the quenching group and emitted in a thermal form, the fluorescence is almost completely quenched, and the fluorescence background is extremely low. When the circular region of the molecular beacon is hybridized with target DNA with completely complementary sequence to form a double-stranded hybrid, the stem region of the molecular beacon is pulled apart, and the distance between the fluorescent group and the quenching group is increased. According to Foerster's theory, the efficiency of central fluorescence energy transfer is inversely proportional to the 6 th power of the distance between the two, and therefore, the fluorescence of the molecular beacon is almost 100% recovered after hybridization, and the detected fluorescence intensity is proportional to the amount of target DNA in solution (FIG. 1). Thus, the ideal molecular beacon is more efficient than the Taqman hydrolysis probe. However, the introduction of a stem region in the molecular beacon, which is not related to the target sequence, often results in some non-specific interaction between the molecular beacon and the template sequence, which leads to an increase in background signal, and thus, affects the detection efficiency. To eliminate this background signal, high requirements are imposed on the design of the molecular beacon, especially on the sequence design of the stem region. In addition, studies have shown that molecular beacons have a good effect for detecting gene mutations (including single-base mismatches, deletions, or insertion mutations) when the sequence of the loop region is short, but in practice, in many cases, the sequence of the loop region is too long due to the low GC content of a specific target sequence region, thereby affecting the detection efficiency. Therefore, it is often difficult to obtain an ideal molecular beacon.
The modification of bases, i.e., the development of artificially simulated non-natural nucleotide pairs, has been carried outIn recent 40 years, 5-methylisocytosine deoxynucleotide-isoguanine deoxynucleotide (iso) and its derivative have been usedMeC-isoG) is classical. The work on the nucleotide pairs in isoC-isoG was first carried out by the American famous synthetic biologist Benner SA, whose team realized the entire central principle of replication, transcription and even translation of isocytosine deoxynucleotide-isoguanine deoxynucleotide (isoC-isoG) artificial expanded nucleic acids in vitro. As shown in FIG. 2, isoC and isoG are isomers of natural nucleotides C and G, respectively, which can perfectly pair themselves but cannot form a pair with natural nucleotides.
In addition to the above manual modification of base structure, there is a large class of non-natural nucleic acids based on modification of base sugar rings, such as Locked Nucleic Acids (LNA). LNA, which broadly refers to an oligonucleotide sequence containing one or more LNA monomers (locked nucleotides), is an artificial mimic nucleic acid that has been rapidly developed in recent years and has been widely used in the fields of molecular diagnostics, gene therapy, and the like. As shown in fig. 3, a methylene bridge is formed between the 2 '-O and 4' -C of the pentose ring of the LNA monomer. LNA does not alter the base pairing of natural nucleic acids, but has greater affinity and greater mismatch recognition relative to natural nucleic acids.
Disclosure of Invention
The invention aims to provide a novel typing detection method and a kit for CASP7 gene rs12415607 polymorphic sites based on molecular beacons of artificial simulated nucleic acids.
In order to achieve the purpose, the invention firstly provides a molecular beacon for detecting rs12415607 site polymorphism of a human CASP7 gene.
The molecular beacon for detecting the rs12415607 site polymorphism of the CASP7 gene consists of a molecular beacon A and a molecular beacon B;
the sequence of the molecular beacon A is a sequence 2 in a sequence table, wherein the 2 nd position of the sequence 2 is a 5-methyl isocytosine deoxynucleotide residue, the 3 rd position is an isoguanine deoxynucleotide residue, the 15 th position is a locked nucleotide residue, the 29 th position is a 5-methyl isocytosine deoxynucleotide residue, the 30 th position is an isoguanine deoxynucleotide residue, and the rest nucleotide residues are natural nucleotide residues;
the sequence of the molecular beacon B is a sequence 3 in a sequence table, wherein the 2 nd position of the sequence 3 is a 5-methyl isocytosine deoxynucleotide residue, the 3 rd position is an isoguanine deoxynucleotide residue, the 15 th position is a locked nucleotide residue, the 29 th position is a 5-methyl isocytosine deoxynucleotide residue, the 30 th position is an isoguanine deoxynucleotide residue, and the rest nucleotide residues are natural nucleotide residues.
The 7 th to 25 th sites of the molecular beacon A and the molecular beacon B are both circular region sequences, and the 1 st to 6 th sites and the 26 th to 31 th sites are both stem region sequences.
The circular regions of the molecular beacon A and the molecular beacon B both target to the rs12415607 locus of the CASP7 gene. Wherein the molecular beacon A targets the 'C' of the rs12415607 locus of the CASP7 gene; the molecular beacon B targets the 'A' of the rs12415607 locus of the CASP7 gene.
Furthermore, two ends of the molecular beacon A and the molecular beacon B are also marked with a fluorescent group and a quenching group, and the fluorescent groups marked by the molecular beacon A and the molecular beacon B are different. The molecular beacon A and the molecular beacon B can be the same or different in labeled quenching group.
In each molecular beacon, the fluorescence emitted by the fluorophore can be absorbed by the quencher. The fluorescent group and the quenching group can be respectively positioned at the 5 'terminal and the 3' terminal of the basic molecular beacon, and the positions of the fluorescent group and the quenching group can be exchanged as long as the requirement that the fluorescence emitted by the fluorescent group in the basic molecular beacon in a free state can be quenched by the quenching group is met.
Further, the fluorophore may be FAM, Hex, TET, Cy3, JOE; the quencher group can be Dabcyl, TAMRA. In the invention, the 5 'end of the molecular beacon A is marked with FAM fluorescent group, and the 3' end is marked with Dabcyl quenching group; the 5 'end of the molecular beacon B is marked with a VIC fluorescent group, and the 3' end is marked with a Dabcyl quenching group.
In order to achieve the aim, the invention further provides a kit for detecting the rs12415607 site polymorphism of the CASP7 gene.
The reagent set for detecting rs12415607 site polymorphism of human CASP7 gene consists of the molecular beacon and a primer pair capable of obtaining a recognition sequence containing the circular region of the molecular beacon by amplification from human genome.
In the above-mentioned kit, the primer pair is composed of a single-stranded DNA represented by sequence 4 in the sequence table and a single-stranded DNA represented by sequence 5 in the sequence table.
In the above kit, the molecular beacon and the primer pair are packaged independently. The molar ratio of the molecular beacon A to the molecular beacon B in the molecular beacon can be 1: 1; the molar ratio of the two single-stranded DNAs in the primer pair may be 1: 1. The molar ratio of the molecular beacon A and the molecular beacon B in the kit to the two single-stranded DNAs of the primer pair can be 2:2:5: 5.
In order to achieve the purpose, the invention also provides a kit for detecting rs12415607 site polymorphism of the CASP7 gene.
The kit for detecting the rs12415607 site polymorphism of the CASP7 gene comprises the molecular beacon or the reagent set.
The kit can also comprise positive quality control, negative quality control and other reagents. The other reagents can be reaction buffer, dNTPs and MgCl2Solution, DNA polymerase and/or nuclease-free water. The positive quality control comprises a recombinant plasmid 1, a recombinant plasmid 2 and a recombinant plasmid 3. The recombinant plasmid 1 is obtained by replacing a DNA fragment between EcoRV and SmaI recognition sequences in an escherichia coli cloning vector pUC57 with a DNA fragment shown in a sequence 1 (the rs12415607 site of a CASP7 gene in the sequence 1 is C); the recombinant plasmid 2 is obtained by replacing a DNA fragment between EcoRV and SmaI recognition sequences in an escherichia coli cloning vector pUC57 with a DNA fragment shown in a sequence 1 (the rs12415607 locus of a CASP7 gene in the sequence 1 is A); the above-mentionedThe recombinant plasmid 3 is obtained by mixing the recombinant plasmid 1 and the recombinant plasmid 2 according to a molar ratio of 1: 1. The negative quality control can be specifically nuclease-free water. The DNA polymerase can be EX Taq DNA polymerase.
In order to achieve the above objects, the present invention also provides a novel use of the above molecular beacon or the above kit.
The invention provides application of the molecular beacon or the reagent set in detecting rs12415607 site polymorphism of a CASP7 gene.
The invention also provides application of the molecular beacon or the kit in predicting or assisting in predicting the risk of the lung cancer of an individual.
In order to achieve the above objects, the present invention finally provides a method for detecting rs12415607 site polymorphism of human CASP7 gene.
The method for detecting the rs12415607 site polymorphism of the CASP7 gene comprises the following steps: and detecting a sample to be detected by using the molecular beacon or the reagent set, and determining the rs12415607 site polymorphism of the CASP7 gene in the sample to be detected according to the change of a fluorescence signal in the sample to be detected.
In the method, the step of detecting the sample to be detected by using the molecular beacon or the kit of reagents is to detect the DNA of the sample to be detected by using the molecular beacon or the kit of reagents.
The method for determining rs12415607 site polymorphism of CASP7 gene in a sample to be detected according to change of a fluorescence signal in the sample to be detected comprises the following steps:
if the sample to be detected releases FAM fluorescence signals, does not release VIC fluorescence signals, and the value of the FAM fluorescence signals is continuously increased, the genotype of the rs12415607 locus of the CASP7 gene of the sample to be detected is or is candidate to be CC genotype;
if the sample to be detected releases the VIC fluorescent signal, does not release the FAM fluorescent signal, and the value of the VIC fluorescent signal is continuously increased, the genotype of the rs12415607 locus of the CASP7 gene of the sample to be detected is or is selected as the AA genotype;
and if the sample to be detected releases the VIC fluorescence signal and the FAM fluorescence signal, and both the FAM fluorescence signal value and the VIC fluorescence signal value are continuously increased, the genotype of the rs12415607 locus of the CASP7 gene of the sample to be detected is or is selected as the AC genotype.
The CC genotype refers to a homozygote of C in basic groups of rs12415607 locus of CASP7 gene on two homologous chromosomes of the DNA of a sample to be detected;
the AA genotype refers to a homozygote of A in the basic groups of rs12415607 sites of CASP7 genes on two homologous chromosomes of the DNA of a sample to be detected;
the AC genotype refers to a heterozygote of bases at rs12415607 sites of CASP7 genes on two homologous chromosomes of the DNA of a sample to be detected, wherein the bases are A and C.
In the above method, the sample to be tested may be a blood sample of a person to be tested.
In the above molecular beacon or kit or application or method, the rs12415607 site of the CASP7 gene is located at the 51 st site of the sequence 1.
Compared with the prior art, the invention has the following beneficial effects: the method for judging the rs12415607 site polymorphism of the CASP7 gene based on the gene specificity PCR combined with the artificial simulated nucleic acid molecular beacon, provided by the invention, has the advantages of high accuracy, high detection speed, simplicity in operation, objective result interpretation, less closed-tube reaction pollution and the like, and is very suitable for large-scale clinical development.
Drawings
Fig. 1 is a schematic diagram of the operation of a molecular beacon.
FIG. 2 is a diagram of the non-natural nucleotide isoguanine nucleotide residue (isoG) and the non-natural nucleotide 5-methylisocytosine deoxynucleotide residue (iso)MeC) The structure of (1).
FIG. 3 is a diagram of the structure of locked nucleotide residues.
FIG. 4 is a schematic diagram of the CC genotype-specific amplification curve at the rs12415607 locus of the human CASP7 gene in example 2 of the present invention.
FIG. 5 is a schematic diagram of the AA genotype specific amplification curve at the rs12415607 site of the human CASP7 gene in example 2 of the present invention.
FIG. 6 is a schematic diagram of the genotype-specific amplification curve of human CASP7 gene rs12415607 site AC in example 2.
FIG. 7 is a schematic diagram of the amplification curve of the standard sample 1 detected using the primer pair SEQ1 and SEQ2, the common Taqman probe SEQ5-FAM and SEQ 6-VIC.
FIG. 8 is a schematic diagram of the amplification curve of the standard sample 2 detected using the primer pair SEQ1 and SEQ2, the common Taqman probe SEQ5-FAM and SEQ 6-VIC.
Sequence listing
<110> Beijing FuanHua Biotech Co., Ltd
<160>7
<170>SIPOSequenceListing 1.0
<210>1
<211>101
<212>DNA
<213> Artificial sequence
<400>1
acgggaaagc aagcaaacag aaagagaaga tggcgttctt gcctggtatc mgaccactaa 60
gcatgtactc cattcacgca aggatttggt ggttcctaaa a 101
<210>2
<211>31
<212>DNA
<213> Artificial sequence
<400>2
ccgacacctg gtatccgacc actaatgtcg g 31
<210>3
<211>31
<212>DNA
<213> Artificial sequence
<400>3
ccgacacctg gtatcagacc actaatgtcg g 31
<210>4
<211>23
<212>DNA
<213> Artificial sequence
<400>4
acgggaaagc aagcaaacag aaa 23
<210>5
<211>24
<212>DNA
<213> Artificial sequence
<400>5
ttttaggaac caccaaatcc ttgc 24
<210>6
<211>24
<212>DNA
<213> Artificial sequence
<400>6
cttgcctggt atccgaccac taag 24
<210>7
<211>24
<212>DNA
<213> Artificial sequence
<400>7
cttgcctggt atcagaccac taag 24
Claims (8)
1. The molecular beacon for detecting rs12415607 locus polymorphism of the CASP7 gene consists of a molecular beacon A and a molecular beacon B;
the sequence of the molecular beacon A is a sequence 2 in a sequence table, wherein the 2 nd position of the sequence 2 is a 5-methyl isocytosine deoxynucleotide residue, the 3 rd position is an isoguanine deoxynucleotide residue, the 15 th position is a locked nucleotide residue, the 29 th position is a 5-methyl isocytosine deoxynucleotide residue, the 30 th position is an isoguanine deoxynucleotide residue, and the rest nucleotide residues are natural nucleotide residues;
the sequence of the molecular beacon B is a sequence 3 in a sequence table, wherein the 2 nd position of the sequence 3 is a 5-methyl isocytosine deoxynucleotide residue, the 3 rd position is an isoguanine deoxynucleotide residue, the 15 th position is a locked nucleotide residue, the 29 th position is a 5-methyl isocytosine deoxynucleotide residue, the 30 th position is an isoguanine deoxynucleotide residue, and the rest nucleotide residues are natural nucleotide residues.
2. The molecular beacon of claim 1, wherein: and fluorescent groups and quenching groups are marked at two ends of the molecular beacon A and the molecular beacon B, and the fluorescent groups marked by the molecular beacon A and the molecular beacon B are different.
3. The molecular beacon of claim 2, wherein: the molecular beacon A is marked with FAM fluorophore; the molecular beacon B is marked with a VIC fluorescent group.
4. A kit for detecting rs12415607 site polymorphism of human CASP7 gene, which comprises the molecular beacon as claimed in any one of claims 1-3 and a primer pair capable of amplifying from human genome to obtain a primer pair containing the recognition sequence of the circular region of the molecular beacon as claimed in any one of claims 1-3.
5. The kit of claim 4, wherein: the primer pair consists of a single-stranded DNA shown in a sequence 4 in a sequence table and a single-stranded DNA shown in a sequence 5 in the sequence table.
6. A kit for detecting rs12415607 site polymorphism of human CASP7 gene, which comprises the molecular beacon of any one of claims 1-3 or the reagent set of claim 4 or 5.
7. Use of the molecular beacon of any one of claims 1 to 3 or the kit of parts of claims 4 or 5 or the kit of parts of claim 6 for detecting the rs12415607 site polymorphism of the CASP7 gene of human;
or, the use of a molecular beacon according to any one of claims 1 to 3 or a kit of parts according to claim 4 or 5 or a kit according to claim 6 for predicting or aiding in the prediction of the risk of an individual to develop lung cancer.
8. The method for detecting the rs12415607 site polymorphism of the CASP7 gene of the human beings comprises the following steps: detecting a sample to be detected by using the molecular beacon as claimed in any one of claims 1 to 3 or the reagent set as claimed in claim 4 or 5, and determining rs12415607 site polymorphism of the CASP7 gene in the sample to be detected according to the change of a fluorescence signal in the sample to be detected.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811598534.3A CN111378746A (en) | 2018-12-27 | 2018-12-27 | Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811598534.3A CN111378746A (en) | 2018-12-27 | 2018-12-27 | Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111378746A true CN111378746A (en) | 2020-07-07 |
Family
ID=71222374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811598534.3A Pending CN111378746A (en) | 2018-12-27 | 2018-12-27 | Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111378746A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1164260A (en) * | 1994-08-30 | 1997-11-05 | 奇龙公司 | Reduction of nonspecific hybridization by using novel base-pairing schemes |
| US20040005595A1 (en) * | 2001-09-28 | 2004-01-08 | Browne Kenneth A. | Inversion probes |
-
2018
- 2018-12-27 CN CN201811598534.3A patent/CN111378746A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1164260A (en) * | 1994-08-30 | 1997-11-05 | 奇龙公司 | Reduction of nonspecific hybridization by using novel base-pairing schemes |
| US20040005595A1 (en) * | 2001-09-28 | 2004-01-08 | Browne Kenneth A. | Inversion probes |
Non-Patent Citations (3)
| Title |
|---|
| PINPIN SHENG等: "Design of a novel molecular beacon: modification of the stem with artificially genetic alphabet", CHEM COMMUN (CAMB), no. 41, pages 1 * |
| 滑峰;周清华;: "肺癌相关单核苷酸多态性的研究进展", 中国肺癌杂志, no. 02, pages 159 * |
| 齐明键: "凋亡相关基因多态性及环境行为因素与噪声性听力损失的关联研究", 中国优秀硕士学位论文全文数据库 医药卫生科技辑, no. 2, pages 2 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108368557B (en) | Isothermal-based bifunctional oligonucleotides and methods for nucleic acid amplification and determination using the same | |
| JP6876437B2 (en) | DNA amplification method based on strand invasion | |
| US10689718B2 (en) | HEV Assay | |
| US20180073087A1 (en) | Qualitative and absolute quantification kit for detecting hepatitis b virus cccdna | |
| CN113046424B (en) | Preparation method of nucleic acid composition for PCR detection, nucleic acid composition prepared by preparation method and PCR detection method | |
| WO2011062258A1 (en) | Primer set for amplification of mthfr gene, mthfr gene amplification reagent comprising same, and use of same | |
| CN107338287B (en) | Kit and method for detecting sheep BMPR-IB gene A746G mutation by Taqman-MGB probe | |
| CN105624315A (en) | Primers and reagent kit for detecting polymorphism of ALDH2 gene c.1510 locus | |
| CN111996244A (en) | Composition for detecting single nucleotide polymorphism and application thereof | |
| CN110885887B (en) | Artificial simulation nucleic acid molecular beacon and kit for detecting rs1517114 site polymorphism of C8orf34 gene | |
| CN111378746A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs12415607 site polymorphism of CASP7 gene | |
| CN110819725B (en) | Method and kit for detecting helicobacter pylori clarithromycin drug-resistant site based on artificial simulation nucleic acid molecular beacon | |
| CN111593122A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting XPC gene rs2228001 site polymorphism | |
| CN111378750A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs885004 site polymorphism of SLC28A3 gene | |
| CN111378749A (en) | Artificial simulation nucleic acid molecular beacon and kit for detecting polymorphism of DYNC2H1 gene rs716274 site | |
| CN111593123A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs2227310 site polymorphism of CASP7 gene | |
| CN105695613A (en) | Method for detecting polymorphism of human gastric cancer susceptible genes IL17A rs3748067 by aid of ApoI | |
| CN111593105A (en) | Artificial simulated nucleic acid molecular beacon and kit for detecting polymorphism of rs4880 site of SOD2 gene | |
| CN111593106A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting polymorphism of rs12041331 locus of PEAR1 gene | |
| CN111575370A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting polymorphism of site rs7853758 of SLC28A3 gene | |
| CN111378748A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs1801019 site polymorphism of UMPS gene | |
| CN111593102A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs10306114 site polymorphism of PTGS1 gene | |
| CN111593101A (en) | Artificial mimic nucleic acid molecular beacon and kit for detecting rs9923231 site polymorphism of VKORC1 gene | |
| CN111593100A (en) | Artificial simulated nucleic acid molecular beacon and kit for detecting GP1BA gene rs6065 site polymorphism | |
| Kitaoka et al. | Simultaneous visual detection of single-nucleotide variations in tuna DNA using DNA/RNA chimeric probes and ribonuclease A |
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 |