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Dot blot is a quick method for detecting biological samples, such as proteins or nucleic acids, using a process similar to western and southern blotting. Instead of transferring from a gel, samples are directly dotted onto a membrane for detection. Due to its simplicity, dot blot is utilized in various applications, including monitoring labeling efficiency with different probes, estimating protein concentration, and comparing antibody performance. For example, 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 the reference standard, starting with a 10-fold dilution to ensure a wide range of samples.
