Determine the labeling efficiency in terms of μg (expected yield of a standard labeling reaction is 20 μg of DIG labeled RNA per μg linearized template DNA after the DIG RNA labeling reaction).
In general, it is possible to use DNA probes for northern blots (PCR or random-primed DNA probes). It is however important to keep in mind that DNA probes are best detecting abundant messages, such as housekeeping genes.
Determine the labeling efficiency in terms of μg (expected yield of a standard labeling reaction is 20 μg of DIG labeled RNA per μg linearized template DNA after the DIG RNA labeling reaction).
In order to specifically detect an antigen or target molecule immobilized on a solid support, unoccupied binding sites on the support must be blocked against binding by probe and detection molecules.
An introduction to both Northern and Southern blotting, popular methods for the transfer of macromolecules to membranous support. This article also offers a Southern blot protocol and a northern blot protocol.
Calculate the melting temperature (Tm) of oligonucleotides using theoretical method like the nearest neighbor or the basic method. The DNA Tm is the temperature at which 50% of the oligonucleotide is duplexed with its perfect complement and 50% is free
Dextran sulfates are supplied as the sodium salt forms, making them soluble and stable in water. Dextran sulfate contains approximately 17% sulfur which is equivalent to approximately 2.3 sulfate groups per glucosyl residue.