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Dot blot is a quick and straightforward method for detecting biological samples such as proteins or nucleic acids. It operates similarly to western and southern blotting, but instead of transferring samples from a gel, samples are directly dotted onto a membrane for detection. Due to its simplicity, dot blot is versatile and used for various applications, including monitoring labeling efficiency with different probes, estimating protein concentrations, and comparing antibody performances. For instance, to determine labeling efficiency of a biotin-labeled sample, one would compare it to a known biotinylated reference standard by making serial dilutions of both samples in the same manner, starting with a 10-fold serial dilution to provide a broad range of concentrations.
