Hybridization and detection of digoxigenin probes on RNA blots.
نویسندگان
چکیده
This article describes nonradioactive probing of a Northern blot. The method employs digoxigenin-labeled probes. Antidigoxigenin antibody/alkaline phosphate conjugate, and a chemiluminescent substrate are subsequently used in the detection system.
منابع مشابه
Non-radioactive labeling of RNA transcripts in vitro with the hapten digoxigenin (DIG); hybridization and ELISA-based detection.
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 perfor...
متن کاملIn situ hybridization with non-radioactive digoxigenin-11-UTP-labeled cRNA probes: localization of developmentally regulated mouse tenascin mRNAs.
An improved method for in situ hybridization was developed in order to identify the tissue-specific expression of messenger RNA (mRNA) for the novel extracellular matrix glycoprotein, tenascin, during mouse development. Non-radioactive RNA probes were generated by incorporating digoxigenin-11-UTP instead of conventional isotopic labels. Hybridization of anti-sense probes to complementary mRNAs ...
متن کاملMultiplexed miRNA northern blots via hybridization chain reaction
Northern blots enable detection of a target RNA of interest in a biological sample using standard benchtop equipment. miRNAs are the most challenging targets as they must be detected with a single short nucleic acid probe. With existing approaches, it is cumbersome to perform multiplexed blots in which several RNAs are detected simultaneously, impeding the study of interacting regulatory elemen...
متن کاملWhole-mount fluorescence in situ hybridization and antibody staining of Drosophila embryos.
Whole-mount RNA in situ hybridizations with digoxigenin-conjugated probes and alkaline phosphatase biochemistry have been used widely for many years to map expression pattern domains in the Drosophila embryo. To capitalize on the number of molecular markers in the central nervous system (CNS) and to enable expression analysis at the single-cell level, fluorescence in situ hybridization procedur...
متن کاملDevelopment of an Alu-PCR Amplified YAC Probe Suitable for Enumeration of Chromosome 13 on Uncultured Lymphocytes and Amniocytes by Fluorescence in situ Hybridization
The main objective of the present study was to develop an efficient and reliable probe to be routinely used for detection of chromosome 13 copy numbers by interphase FISH. To achieve this, a Yeast Artificial Chromosome (YAC) containing sequences specific for human 13q12 (744D11), was cultured and the whole yeast genomic DNA was extracted. The human insert within the isolated DNA was amplified b...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Molecular biotechnology
دوره 6 1 شماره
صفحات -
تاریخ انتشار 1994