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Dot blot is a quick method for detecting biological samples such as proteins or nucleic acids, similar to western and southern blotting but without transferring samples from a gel. Instead, samples are dotted directly onto a membrane for detection. Due to its simplicity, dot blot is used in various applications, including monitoring labeling efficiency with different probes, estimating protein concentration in samples, and comparing antibody performance. For instance, to assess labeling efficiency with biotin, one would compare a labeled sample to a known biotinylated reference. This involves making serial dilutions of both the test sample and the reference standard, typically starting with a 10-fold dilution for a broad range of samples.
