JP4069133B2 - 生物試料に含まれる目的遺伝子を増幅する方法 - Google Patents
生物試料に含まれる目的遺伝子を増幅する方法 Download PDFInfo
- Publication number
- JP4069133B2 JP4069133B2 JP2005327174A JP2005327174A JP4069133B2 JP 4069133 B2 JP4069133 B2 JP 4069133B2 JP 2005327174 A JP2005327174 A JP 2005327174A JP 2005327174 A JP2005327174 A JP 2005327174A JP 4069133 B2 JP4069133 B2 JP 4069133B2
- Authority
- JP
- Japan
- Prior art keywords
- reaction
- pcr
- primer
- concentration
- oligo
- 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.)
- Expired - Fee Related
Links
- 108090000623 proteins and genes Proteins 0.000 title claims description 84
- 238000000034 method Methods 0.000 title claims description 70
- 239000012472 biological sample Substances 0.000 title claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 155
- 108020004414 DNA Proteins 0.000 claims description 46
- 102000053602 DNA Human genes 0.000 claims description 44
- 238000010839 reverse transcription Methods 0.000 claims description 37
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 claims description 24
- 210000003719 b-lymphocyte Anatomy 0.000 claims description 23
- 230000006820 DNA synthesis Effects 0.000 claims description 22
- 238000007857 nested PCR Methods 0.000 claims description 21
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 claims description 15
- 239000000758 substrate Substances 0.000 claims description 14
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 claims description 13
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 claims description 13
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 claims description 13
- 238000000746 purification Methods 0.000 claims description 13
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 12
- 108010083359 Antigen Receptors Proteins 0.000 claims description 11
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 claims description 11
- 229910001425 magnesium ion Inorganic materials 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 9
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 238000000926 separation method Methods 0.000 claims description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 6
- 238000003786 synthesis reaction Methods 0.000 claims description 6
- 108020004682 Single-Stranded DNA Proteins 0.000 claims description 5
- -1 that Chemical compound 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 32
- 230000003321 amplification Effects 0.000 description 25
- 238000003199 nucleic acid amplification method Methods 0.000 description 25
- 239000000047 product Substances 0.000 description 24
- 238000001962 electrophoresis Methods 0.000 description 15
- 102100033215 DNA nucleotidylexotransferase Human genes 0.000 description 13
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 13
- 210000004698 lymphocyte Anatomy 0.000 description 11
- 102100034343 Integrase Human genes 0.000 description 10
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 10
- 238000007796 conventional method Methods 0.000 description 8
- 108020004999 messenger RNA Proteins 0.000 description 8
- 108700042075 T-Cell Receptor Genes Proteins 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 239000011543 agarose gel Substances 0.000 description 6
- 102000006382 Ribonucleases Human genes 0.000 description 5
- 108010083644 Ribonucleases Proteins 0.000 description 5
- 102100022887 GTP-binding nuclear protein Ran Human genes 0.000 description 4
- 101000774835 Heteractis crispa PI-stichotoxin-Hcr2o Proteins 0.000 description 4
- 101000620756 Homo sapiens GTP-binding nuclear protein Ran Proteins 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 229910021645 metal ion Inorganic materials 0.000 description 4
- 239000013600 plasmid vector Substances 0.000 description 4
- 101100393821 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) GSP2 gene Proteins 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 210000001671 embryonic stem cell Anatomy 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000011777 magnesium Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 229920001917 Ficoll Polymers 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000003161 ribonuclease inhibitor Substances 0.000 description 2
- 239000003419 rna directed dna polymerase inhibitor Substances 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 230000000087 stabilizing effect Effects 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 102000006306 Antigen Receptors Human genes 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 102000028180 Glycophorins Human genes 0.000 description 1
- 108091005250 Glycophorins Proteins 0.000 description 1
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 1
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 101000934346 Homo sapiens T-cell surface antigen CD2 Proteins 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 108091008874 T cell receptors Proteins 0.000 description 1
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 1
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- ZFTFAPZRGNKQPU-UHFFFAOYSA-N dicarbonic acid Chemical compound OC(=O)OC(O)=O ZFTFAPZRGNKQPU-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 238000007898 magnetic cell sorting Methods 0.000 description 1
- 239000011325 microbead Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005327174A JP4069133B2 (ja) | 2004-11-19 | 2005-11-11 | 生物試料に含まれる目的遺伝子を増幅する方法 |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2004336090 | 2004-11-19 | ||
| JP2005327174A JP4069133B2 (ja) | 2004-11-19 | 2005-11-11 | 生物試料に含まれる目的遺伝子を増幅する方法 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2006166907A JP2006166907A (ja) | 2006-06-29 |
| JP2006166907A5 JP2006166907A5 (enExample) | 2007-03-22 |
| JP4069133B2 true JP4069133B2 (ja) | 2008-04-02 |
Family
ID=36668269
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2005327174A Expired - Fee Related JP4069133B2 (ja) | 2004-11-19 | 2005-11-11 | 生物試料に含まれる目的遺伝子を増幅する方法 |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4069133B2 (enExample) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10533204B2 (en) | 2014-05-30 | 2020-01-14 | National University Corporatlon University of Toyama | Method for amplifying a T cell receptor (TCR) cDNA |
| EP4019648A4 (en) * | 2019-08-20 | 2023-10-04 | Director-General of National Institute of Infectious Diseases | Method for amplifying nucleotide sequence, and sequence determination method |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009098988A1 (ja) * | 2008-02-04 | 2009-08-13 | Genecare Research Institute Co., Ltd. | siRNA検出方法 |
| JP6164759B2 (ja) | 2013-11-21 | 2017-07-19 | Ｒｅｐｅｒｔｏｉｒｅ Ｇｅｎｅｓｉｓ株式会社 | Ｔ細胞受容体およびｂ細胞受容体レパトアの解析システムならびにその治療および診断への利用 |
| CN109517889B (zh) | 2017-09-18 | 2022-04-05 | 苏州吉赛基因测序科技有限公司 | 一种基于高通量测序分析寡核苷酸序列杂质的方法及应用 |
| CN113774496A (zh) * | 2021-10-11 | 2021-12-10 | 湖南大地同年生物科技有限公司 | 一种液相捕获文库构建方法 |
-
2005
- 2005-11-11 JP JP2005327174A patent/JP4069133B2/ja not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10533204B2 (en) | 2014-05-30 | 2020-01-14 | National University Corporatlon University of Toyama | Method for amplifying a T cell receptor (TCR) cDNA |
| EP4019648A4 (en) * | 2019-08-20 | 2023-10-04 | Director-General of National Institute of Infectious Diseases | Method for amplifying nucleotide sequence, and sequence determination method |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2006166907A (ja) | 2006-06-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11898209B2 (en) | Digital analysis of circulating tumor cells in blood samples | |
| US5681726A (en) | Method of double stranded DNA synthesis | |
| EP3485043A1 (en) | Single-cell transcript sequencing | |
| JP2009284834A (ja) | 大規模並列核酸分析方法 | |
| JP4069133B2 (ja) | 生物試料に含まれる目的遺伝子を増幅する方法 | |
| US20220259653A1 (en) | Dimer avoided multiplex polymerase chain reaction for amplification of multiple targets | |
| US9708603B2 (en) | Method for amplifying cDNA derived from trace amount of sample | |
| Shcheglov et al. | Normalization of cDNA libraries | |
| EP1854882A1 (en) | Method for obtaining subtraction polynucleotide | |
| WO2006054492A1 (ja) | 生物試料に含まれる目的遺伝子を増幅する方法 | |
| Weiss et al. | Optimizing utilization of DNA from rare or archival anthropological samples | |
| Hubank et al. | Cloning of apoptosis-related genes by representational difference analysis of cDNA | |
| Warren | Single-cell gene-expression analysis by quantitative RT-PCR | |
| Payet et al. | Small RNA Profiling by Next-Generation Sequencing Using High-Definition Adapters | |
| Klebes et al. | Linear RNA amplification for the production of microarray hybridization probes | |
| JP2025181919A (ja) | Ｒｎａオリゴヌクレオチドをシークエンシングする方法 | |
| JP4300047B2 (ja) | 核酸の合成方法 | |
| WO2012150455A1 (en) | Sensitive preparation and amplification of eukaryotic derived nucleic acid for transcriptional profiling | |
| CA3151485A1 (en) | A multiplex method of preparing a sequencing library | |
| EP1521829A2 (en) | Target dna | |
| Schatz | Representational difference analysis of cDNA | |
| SIRARD et al. | Gene Subtraction and Analysis | |
| Mastrangeli et al. | Cloning of Homologous Genes by Gene-Capture PCR |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070206 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20070206 |
|
| A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20070206 |
|
| A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20070404 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070508 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070709 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20071009 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20071016 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20080108 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20080111 |
|
| FPAY | Renewal fee payment (prs date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110118 Year of fee payment: 3 |
|
| R150 | Certificate of patent (=grant) or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| LAPS | Cancellation because of no payment of annual fees |