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Dot blot is a straightforward method for detecting biological samples, such as proteins or nucleic acids. Unlike western and southern blotting, samples are directly dotted onto a membrane rather than transferred from a gel. Its simplicity allows for various applications, including monitoring labeling efficiency with different probes, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency with a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and the reference, ensuring they are diluted identically; a 10-fold serial dilution is effective for providing a broad range of samples.
