JPH09187299A - Primer for pcr - Google Patents
Primer for pcrInfo
- Publication number
- JPH09187299A JPH09187299A JP8027222A JP2722296A JPH09187299A JP H09187299 A JPH09187299 A JP H09187299A JP 8027222 A JP8027222 A JP 8027222A JP 2722296 A JP2722296 A JP 2722296A JP H09187299 A JPH09187299 A JP H09187299A
- Authority
- JP
- Japan
- Prior art keywords
- mrna
- pcr
- primer
- human proteins
- tsh
- 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
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、ヒトの細胞に含まれる
ｍＲＮＡの存在確認および抽出に関する。TECHNICAL FIELD The present invention relates to confirmation and extraction of mRNA contained in human cells.
【０００２】[0002]
【従来の技術】本発明に関するプライマーは、他に存在
せず、かつ本プライマーがとらえ得る遺伝子産物まで
は、従来の方法では達していない。2. Description of the Related Art There is no other primer relating to the present invention, and a gene product which can be captured by this primer has not been reached by conventional methods.
【０００３】[0003]
【発明が解決しようとする課題】従来のＰＣＲ用プライ
マーでは、理論上は一分子のｍＲＮＡをもとらえ得る
が、実際上では一細胞内の一分子のｍＲＮＡをとらえる
ことは、きわめて困難であった。With the conventional PCR primers, it is theoretically possible to capture one molecule of mRNA, but in reality, it is extremely difficult to capture one molecule of mRNA in one cell. .
【０００４】[0004]
【課題を解決するための手段】上記目的を達するため
に、本発明のプライマーにおいては、遺伝子産物の内部
に順次プライマーを新たに設計し、遺伝子産物の増輻を
改善したものである。In order to achieve the above object, in the primer of the present invention, a primer is newly designed in sequence inside the gene product to improve the gene product enhancement.
【０００５】[0005]
【作用】複数のプライマーを順次組合せることにより、
目的とする遺伝子産物を増幅する事ができ、検出しよう
とする極微量のｍＲＮＡの存在を容易に知ることができ
る。[Operation] By combining multiple primers sequentially,
The target gene product can be amplified, and the presence of a trace amount of mRNA to be detected can be easily known.
【０００６】[0006]
【実施例】甲状腺刺激ホルモンの例について説明する
と、一個の細胞より抽出したｍＲＮＡより、通常の方法
でＴＳＨ−５とＴＳＨ−２を１回目のプライマーとして
ＰＣＲを行ってもＰＣＲ遺伝子産物をとらえることがで
きない。次に２回目のプライマーとして、ＴＳＨ−５と
ＴＳＨ−６を、３回目としてＴＳＨ−１とＴＳＨ−６
を、４回目として、ＴＳＨ−１とＴＳＨ−４もしくは、
ＴＳＨ−３とＴＳＨ−６を組合せてＰＣＲを実施する
が、これでも多くの場合、目的とするＰＣＲ産物を得る
ことができない。最後にＴＳＨ−７とＴＳＨ−４をプラ
イマーとしてＰＣＲを行うと一連のＰＣＲの結果とし
て、大きさが１０６塩基対（表１）のＰＣＲ産物を得る
ことができる。他の蛋白質のｍＲＮＡについても同様
に、前回のＰＣＲ産物の内側にプライマーを設定するこ
とを繰り返すことにより、一個の細胞中の極めてわずか
に存在するｍＲＮＡをとらえることができる。[Examples] An example of thyroid-stimulating hormone will be explained. From the mRNA extracted from one cell, the PCR gene product can be detected even if PCR is carried out by the usual method using TSH-5 and TSH-2 as the first primer. I can't. Next, TSH-5 and TSH-6 were used as the second primer, and TSH-1 and TSH-6 were used as the third primer.
As the fourth time, TSH-1 and TSH-4, or
Although PCR is carried out by combining TSH-3 and TSH-6, in most cases, the desired PCR product cannot be obtained. Finally, when PCR is performed using TSH-7 and TSH-4 as a primer, a PCR product having a size of 106 base pairs (Table 1) can be obtained as a result of a series of PCRs. Similarly, for mRNAs of other proteins, it is possible to capture extremely small amounts of mRNAs in one cell by repeatedly setting primers inside the previous PCR product.
【０００７】[0007]
【発明の効果】本発明は１１種の蛋白質に関するｍＲＮ
Ａについて上記に説明したように構成されているので、
表１に示した遺伝子産物を検出し得る。このことで、そ
れぞれの蛋白質を規定するｍＲＮＡの存在を、一細胞の
中でもとらえることができる。INDUSTRIAL APPLICABILITY The present invention relates to mRN relating to 11 kinds of proteins.
Since it is configured as described above for A,
The gene products shown in Table 1 can be detected. As a result, the presence of mRNA that defines each protein can be detected in one cell.
【０００８】[0008]
【表１】 [Table 1]
Claims (1)
センジャーＲＮＡ（以下ｍＲＮＡ）の存在を確認するた
めのＰＣＣＲ（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒ
ｅａｃｔｉｏｎ）法に用いるプライマー 1. A PCCR (polymerase chain r) for confirming the presence of messenger RNA (hereinafter referred to as mRNA) encoding various human proteins.
primer used in the reaction method
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8027222A JPH09187299A (en) | 1996-01-05 | 1996-01-05 | Primer for pcr |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8027222A JPH09187299A (en) | 1996-01-05 | 1996-01-05 | Primer for pcr |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH09187299A true JPH09187299A (en) | 1997-07-22 |
Family
ID=12215081
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP8027222A Pending JPH09187299A (en) | 1996-01-05 | 1996-01-05 | Primer for pcr |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH09187299A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000040749A2 (en) * | 1999-01-06 | 2000-07-13 | Genenews Inc. | Method for the detection of gene transcripts in blood and uses thereof |
| KR100458505B1 (en) * | 1998-10-07 | 2005-02-04 | (주)바이오니아 | Multiplex primer for non-specific amplification of nucleic acid |
| US7473528B2 (en) | 1999-01-06 | 2009-01-06 | Genenews Inc. | Method for the detection of Chagas disease related gene transcripts in blood |
-
1996
- 1996-01-05 JP JP8027222A patent/JPH09187299A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100458505B1 (en) * | 1998-10-07 | 2005-02-04 | (주)바이오니아 | Multiplex primer for non-specific amplification of nucleic acid |
| WO2000040749A2 (en) * | 1999-01-06 | 2000-07-13 | Genenews Inc. | Method for the detection of gene transcripts in blood and uses thereof |
| WO2000040749A3 (en) * | 1999-01-06 | 2001-07-19 | Genenews Inc | Method for the detection of gene transcripts in blood and uses thereof |
| US7473528B2 (en) | 1999-01-06 | 2009-01-06 | Genenews Inc. | Method for the detection of Chagas disease related gene transcripts in blood |
| US7662558B2 (en) | 1999-01-06 | 2010-02-16 | Genenews Corporation | Method of profiling gene expression in a human subject |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6849404B2 (en) | Polymerase chain reaction of DNA of which base sequence is completely unidentified | |
| Tosar et al. | Mining of public sequencing databases supports a non-dietary origin for putative foreign miRNAs: underestimated effects of contamination in NGS | |
| Gansauge et al. | Selective enrichment of damaged DNA molecules for ancient genome sequencing | |
| CN102912036B (en) | Quickly detection and the method for qualification inanimate object | |
| JP3082942B2 (en) | Labeling and tracking of substances with nucleic acids | |
| EP3470529A1 (en) | Nucleic acid isothermal self-amplification method | |
| ATE188743T1 (en) | OLIGONUCLEOTIDES COMPLEMENTARY TO MYCOBACTERIAL NUCLEIC ACID | |
| JP2014204730A5 (en) | ||
| Raabe et al. | The rocks and shallows of deep RNA sequencing: Examples in the Vibrio cholerae RNome | |
| CN104946629A (en) | Method for fragmenting trace DNA sample and method for establishing DNA library by utilizing trace DNA sample | |
| EP2690180B1 (en) | Method for enhancing efficiency and sensitivity in nucleic acid amplification from biological materials using ionic liquids | |
| US20210403904A1 (en) | Methods for haplotyping with short read sequence technology | |
| WO2016049638A1 (en) | Methods and systems for detection of a genetic mutation | |
| CN113308526A (en) | Fusion primer direct amplification method human mitochondrial whole genome high-throughput sequencing kit | |
| JPH09187299A (en) | Primer for pcr | |
| EP2947156A1 (en) | Optimization of sequencing reactions | |
| EP2510114B1 (en) | Rna analytics method | |
| JP4531888B2 (en) | Method for separating nucleic acids | |
| EP1471035A3 (en) | A carrier of a diamond fine particle for immobilizing virus | |
| CN110777211A (en) | Composite amplification system based on Y-STR locus and Y-indel locus and primer combination used by same | |
| ATE40379T1 (en) | PROCESSES FOR THE DISSOLUTION OF DEOXYRIBONUCLEIC ACID IN NON-AQUEOUS SOLUTIONS AND MIXED SYSTEMS OF NON-AQUEOUS/AQUEOUS SOLUTIONS, THE SOLUTIONS SO OBTAINED AND THEIR USE. | |
| WO2018218136A1 (en) | Reverse complement adapters for the mitigation of umi hopping | |
| AU2021330837A1 (en) | Enhanced sequencing following random DNA ligation and repeat element amplification | |
| CN103547681B (en) | Method for capturing target region and method and system for processing bioinformatics thereof | |
| Bonot et al. | Monitoring the dissemination of the broad-host-range plasmid pB10 in sediment microcosms by quantitative PCR |