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CDNA LIBRARY CONSTRUCTION FROM SMALL AMOUNTS OF UNFRACTIONATED RNA - ASSOCIATION OF CDNA SYNTHESIS WITH POLYMERASE CHAIN-REACTION AMPLIFICATION
被引:21
作者
:
DOMEC, C
论文数:
0
引用数:
0
h-index:
0
机构:
Groupe d'Etude de l'Expression Génétique du Système Nerveux, Institut de Biochimie Cellulaire et Neurochimie, CNRS, 33077 Bordeaux Cedex
DOMEC, C
GARBAY, B
论文数:
0
引用数:
0
h-index:
0
机构:
Groupe d'Etude de l'Expression Génétique du Système Nerveux, Institut de Biochimie Cellulaire et Neurochimie, CNRS, 33077 Bordeaux Cedex
GARBAY, B
FOURNIER, M
论文数:
0
引用数:
0
h-index:
0
机构:
Groupe d'Etude de l'Expression Génétique du Système Nerveux, Institut de Biochimie Cellulaire et Neurochimie, CNRS, 33077 Bordeaux Cedex
FOURNIER, M
BONNET, J
论文数:
0
引用数:
0
h-index:
0
机构:
Groupe d'Etude de l'Expression Génétique du Système Nerveux, Institut de Biochimie Cellulaire et Neurochimie, CNRS, 33077 Bordeaux Cedex
BONNET, J
机构
:
[1]
Groupe d'Etude de l'Expression Génétique du Système Nerveux, Institut de Biochimie Cellulaire et Neurochimie, CNRS, 33077 Bordeaux Cedex
来源
:
ANALYTICAL BIOCHEMISTRY
|
1990年
/ 188卷
/ 02期
关键词
:
D O I
:
10.1016/0003-2697(90)90630-R
中图分类号
:
Q5 [生物化学];
学科分类号
:
071010 ;
081704 ;
摘要
:
We describe here a general and simple procedure for cDNA library construction making use of in vitro amplification of cDNA by polymerase chain reaction (PCR). The first-strand cDNA is synthesized from total RNA with a primer EcoRI-(dT)17 and oligo(dG) tailed. An oligonucleotide, EcoRI-BamHI-(dC)13, is used to prime the second-strand synthesis by the thermostable DNA polymerase of Thermus aquaticus. The doublestranded cDNA is then amplified directly by PCR. A study of the effect of the elongation time on the PCR products showed that a long extension time is necessary to overcome the size heterogeneity of the cDNA population. Starting from 1 μg of total brain RNA, the products obtained ranged from 200 to more than 2000 bp. The presence of the myelin basic protein cDNA sequence was determined. A λgt10 library containing 2 × 106 clones was estblished with the amplified cDNA. No sequences originating from rRNA were detected by Southern blot analysis. The ability to produce representative cDNA libraries from minute amounts of total RNA by this protocol should have many applications to studies of gene expression in small amounts of tissues or cells. © 1990.
引用
收藏
页码:422 / 426
页数:5
相关论文
共 27 条
[1]
PCR-BASED CDNA LIBRARY CONSTRUCTION - GENERAL CDNA LIBRARIES AT THE LEVEL OF A FEW CELLS
BELYAVSKY, A
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
BELYAVSKY, A
VINOGRADOVA, T
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
VINOGRADOVA, T
RAJEWSKY, K
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
RAJEWSKY, K
[J].
NUCLEIC ACIDS RESEARCH,
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CHEHAB, FF
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
CHEHAB, FF
DOHERTY, M
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
DOHERTY, M
CAI, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
CAI, S
KAN, YW
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0
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0
h-index:
0
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UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
KAN, YW
COOPER, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
COOPER, S
RUBIN, EM
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
RUBIN, EM
[J].
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CHELLY, J
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHELLY, J
KAPLAN, JC
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
KAPLAN, JC
MAIRE, P
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
MAIRE, P
GAUTRON, S
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
GAUTRON, S
KAHN, A
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
KAHN, A
[J].
NATURE,
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ALTERNATIVE SPLICING ACCOUNTS FOR THE 4 FORMS OF MYELIN BASIC-PROTEIN
DEFERRA, F
论文数:
0
引用数:
0
h-index:
0
DEFERRA, F
ENGH, H
论文数:
0
引用数:
0
h-index:
0
ENGH, H
HUDSON, L
论文数:
0
引用数:
0
h-index:
0
HUDSON, L
KAMHOLZ, J
论文数:
0
引用数:
0
h-index:
0
KAMHOLZ, J
PUCKETT, C
论文数:
0
引用数:
0
h-index:
0
PUCKETT, C
MOLINEAUX, S
论文数:
0
引用数:
0
h-index:
0
MOLINEAUX, S
LAZZARINI, RA
论文数:
0
引用数:
0
h-index:
0
LAZZARINI, RA
[J].
CELL,
1985,
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RAPID AMPLIFICATION OF COMPLEMENTARY-DNA FROM SMALL AMOUNTS OF UNFRACTIONATED RNA
DOHERTY, PJ
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
DOHERTY, PJ
HUESCACONTRERAS, M
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
HUESCACONTRERAS, M
DOSCH, HM
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
DOSCH, HM
PAN, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
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DULAU, L
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0
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0
DULAU, L
CHEYROU, A
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0
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CHEYROU, A
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DUTILH, B
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0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
DUTILH, B
BEBEAR, C
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0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
BEBEAR, C
RODRIGUEZ, P
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0
引用数:
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机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
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VEKRIS, A
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CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
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BONNET, J
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CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
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GARRET, M
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0
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h-index:
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CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
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←
1
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3
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共 27 条
[1]
PCR-BASED CDNA LIBRARY CONSTRUCTION - GENERAL CDNA LIBRARIES AT THE LEVEL OF A FEW CELLS
BELYAVSKY, A
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
BELYAVSKY, A
VINOGRADOVA, T
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
VINOGRADOVA, T
RAJEWSKY, K
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
UNIV COLOGNE,INST GENET,D-5000 WEYERTAL 121,FED REP GER
RAJEWSKY, K
[J].
NUCLEIC ACIDS RESEARCH,
1989,
17
(08)
: 2919
-
2932
[2]
DETECTION OF SICKLE-CELL-ANEMIA AND THALASSEMIAS
CHEHAB, FF
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
CHEHAB, FF
DOHERTY, M
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
DOHERTY, M
CAI, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
CAI, S
KAN, YW
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
KAN, YW
COOPER, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
COOPER, S
RUBIN, EM
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV CALIF SAN FRANCISCO, DEPT PEDIAT, SAN FRANCISCO, CA 94143 USA
RUBIN, EM
[J].
NATURE,
1987,
329
(6137)
: 293
-
294
[3]
TRANSCRIPTION OF THE DYSTROPHIN GENE IN HUMAN-MUSCLE AND NON-MUSCLE TISSUES
CHELLY, J
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHELLY, J
KAPLAN, JC
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
KAPLAN, JC
MAIRE, P
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
MAIRE, P
GAUTRON, S
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
GAUTRON, S
KAHN, A
论文数:
0
引用数:
0
h-index:
0
机构:
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
CHU COCHIN,BIOCHIM GENET LAB,F-75014 PARIS,FRANCE
KAHN, A
[J].
NATURE,
1988,
333
(6176)
: 858
-
860
[4]
ALTERNATIVE SPLICING ACCOUNTS FOR THE 4 FORMS OF MYELIN BASIC-PROTEIN
DEFERRA, F
论文数:
0
引用数:
0
h-index:
0
DEFERRA, F
ENGH, H
论文数:
0
引用数:
0
h-index:
0
ENGH, H
HUDSON, L
论文数:
0
引用数:
0
h-index:
0
HUDSON, L
KAMHOLZ, J
论文数:
0
引用数:
0
h-index:
0
KAMHOLZ, J
PUCKETT, C
论文数:
0
引用数:
0
h-index:
0
PUCKETT, C
MOLINEAUX, S
论文数:
0
引用数:
0
h-index:
0
MOLINEAUX, S
LAZZARINI, RA
论文数:
0
引用数:
0
h-index:
0
LAZZARINI, RA
[J].
CELL,
1985,
43
(03)
: 721
-
727
[5]
RAPID AMPLIFICATION OF COMPLEMENTARY-DNA FROM SMALL AMOUNTS OF UNFRACTIONATED RNA
DOHERTY, PJ
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
DOHERTY, PJ
HUESCACONTRERAS, M
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
HUESCACONTRERAS, M
DOSCH, HM
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
DOSCH, HM
PAN, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT PATHOL,PISCATAWAY,NJ 08854
PAN, S
[J].
ANALYTICAL BIOCHEMISTRY,
1989,
177
(01)
: 7
-
10
[6]
DIRECTED MUTAGENESIS USING PCR
DULAU, L
论文数:
0
引用数:
0
h-index:
0
DULAU, L
CHEYROU, A
论文数:
0
引用数:
0
h-index:
0
CHEYROU, A
AIGLE, M
论文数:
0
引用数:
0
h-index:
0
AIGLE, M
[J].
NUCLEIC ACIDS RESEARCH,
1989,
17
(07)
: 2873
-
2873
[7]
SPECIFIC AMPLIFICATION OF A DNA-SEQUENCE COMMON TO ALL CHLAMYDIA-TRACHOMATIS SEROVARS USING THE POLYMERASE CHAIN-REACTION
DUTILH, B
论文数:
0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
DUTILH, B
BEBEAR, C
论文数:
0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
BEBEAR, C
RODRIGUEZ, P
论文数:
0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
RODRIGUEZ, P
VEKRIS, A
论文数:
0
引用数:
0
h-index:
0
机构:
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
CNRS, INST BIOCHIM CELLULAIRE & NEUROCHIM, F-33077 BORDEAUX, FRANCE
VEKRIS, A
BONNET, J
论文数:
0
引用数:
0
h-index:
0
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