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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids, operating on principles similar to western and southern blotting, but with samples dotted directly onto a membrane rather than blotted from a gel. Its simplicity allows for diverse applications, including monitoring labeling efficiency with various probes, estimating specific protein concentrations, and comparing antibody performance. For instance, to determine labeling efficiency with a biotin-labeled sample, one would compare it to a known biotinylated reference. This involves creating serial dilutions of both the test sample and reference standard in the same way, with a 10-fold serial dilution providing a wide range of samples for analysis.
