RNA, which complements a specific mRNA or DNA, is generally used to study virus genes, distribution of specific RNA in tissues and cells, integration of viral DNA into genomes, transcription, etc. Compared with DNA probes which are preferred for detection of the presence of DNA/RNA from specific species or subspecies, RNA probes are commonly for genetic studies. RNA probes can be subdivided by category into plus-sense RNA probes, minus-sense RNA probes, and antisense RNA probes. RNA probes can be used for Northern blotting, RNase protection assays, Southern blotting, downstream of polymerase chain reaction (PCR), and in situ hybridization analysis.
Northern blotting
A Northern blot is a laboratory method used to detect specific RNA molecules. The RNA under study is fractionated by gel electrophoresis. The molecules are then transferred to a membrane that is incubated with the labeled probe(s). Hybridization of complementary sequences allows visualization of target RNA sequences.
RNase protection assays
RPA also depends on the hybridization of probes to target RNAs, but the hybridization takes place in solution. RNA Probes are designed to have non-hybridizing regions. Controlled RNase digestion of the hybrids followed by separation and detection of the remaining probes allows for quantitative analysis of the target RNA in a sample and can provide information on target RNA structure.
In situ hybridization
ISH experiments are conducted to localize RNA or DNA targets in cells and tissues. This technique involves cultured cells or tissue sections as the substrates for hybridization and detection. Cells or tissues are processed so that their endogenous nucleic acids are fixed in place, but available for hybridization to and detection by labeled probes.
Southern blotting
Southern blotting is a technique for transfer of DNA molecules from an electrophoresis gel to a nitrocellulose or nylon membrane, and is carried out prior to detection of specific molecules by hybridization probing. This technique involves the fractionation and transfer of DNA to membranes. Membranes are then are incubated with the labeled probe. Hybridization of complementary sequences allows visualization of target DNA sequences.
The RNA Probe Kit
The RNA Probe Kit allows the rapid and convenient recovery of RNA from any RNA labeling reaction. This kit can also be used for general RNA clean up, where other methods may lead to nuclease contamination or loss of samples. Binding RNA spin-columns and optimized buffers facilitate RNA isolation free of nucleotides, unincorporated labels, enzymes and salts.
