NONRADIOACTIVE LABELING OF RNA TRANSCRIPTS INVITRO WITH THE HAPTEN DIGOXIGENIN (DIG) - HYBRIDIZATION AND ELISA-BASED DETECTION

被引：179
作者：
HOLTKE, HJ
KESSLER, C
机构：
[1] Boehringer Mannheim GmbH, Biochemical Research Center, D-81 22 Penzberg
来源：
NUCLEIC ACIDS RESEARCH | 1990年 / 18卷 / 19期
关键词：
D O I：
10.1093/nar/18.19.5843
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We have developed a system for the enzymatic in vitro synthesis of non-radioactively labeled RNA which is derivatized with the hapten digoxigenin (DIG). The labeling reaction as well as the conditions for hybridization and detection of hybrids by an antibody-conjugate and a coupled colour reaction were analyzed and adapted for high sensitivity and low background. In addition, data on the performance and sensitivity of digoxigenin-labeled RNA probes in Southern and Northern blots are presented. © 1990 Oxford University Press.
引用
收藏
页码：5843 / 5851
页数：9
相关论文
共 44 条
[1]  
Ausubel FM., 1995, MOL REPROD DEV, V3rd edn, DOI DOI 10.1002/MRD.1080010210
[2]   CHEMI-LUMINESCENT DETECTION OF DNA - APPLICATION FOR DNA SEQUENCING AND HYBRIDIZATION [J].
BECK, S ;
OKEEFFE, T ;
COULL, JM ;
KOSTER, H .
NUCLEIC ACIDS RESEARCH, 1989, 17 (13) :5115-5123
[3]   1,2-DIOXETANES - NOVEL CHEMI-LUMINESCENT ENZYME SUBSTRATES - APPLICATIONS TO IMMUNOASSAYS [J].
BRONSTEIN, I ;
EDWARDS, B ;
VOYTA, JC .
JOURNAL OF BIOLUMINESCENCE AND CHEMILUMINESCENCE, 1989, 4 (01) :99-111
[4]  
BROWN JE, 1984, NUCLEIC ACIDS RES, V14, P3521
[5]  
BUTLER ET, 1982, J BIOL CHEM, V257, P5772
[6]   COMPARISON OF COLORIMETRIC, FLUORESCENT, AND ENZYMATIC AMPLIFICATION SUBSTRATE SYSTEMS IN AN ENZYME-IMMUNOASSAY FOR DETECTION OF DNA-RNA HYBRIDS [J].
COUTLEE, F ;
VISCIDI, RP ;
YOLKEN, RH .
JOURNAL OF CLINICAL MICROBIOLOGY, 1989, 27 (05) :1002-1007
[7]   DETECTION OF MESSENGER-RNAS IN SEA-URCHIN EMBRYOS BY INSITU HYBRIDIZATION USING ASYMMETRIC RNA PROBES [J].
COX, KH ;
DELEON, DV ;
ANGERER, LM ;
ANGERER, RC .
DEVELOPMENTAL BIOLOGY, 1984, 101 (02) :485-502
[8]   NON-RADIOACTIVE HYBRIDIZATION PROBES PREPARED BY THE CHEMICAL LABELING OF DNA AND RNA WITH A NOVEL REAGENT, PHOTOBIOTIN [J].
FORSTER, AC ;
MCINNES, JL ;
SKINGLE, DC ;
SYMONS, RH .
NUCLEIC ACIDS RESEARCH, 1985, 13 (03) :745-761
[9]  
HAMES BD, 1984, NUCLEIC ACID HYBRIDI
[10]  
HEILES HBJ, 1988, BIOTECHNIQUES, V6, P978
12345