Slot blot

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A slot blot is a membrane to which an array of DNA slots has been blotted using a slot blot manifold which uses a vacuum to filter the water out of the wells leaving only the DNA (cDNA, PCR amplicons, long oligos). A dot blot is the same just that the wells are round and smaller allowing more probes. This is basically an alternative (or a precursor) to microarrays. A radioactive sample can be hybridized to it allowing the researcher to detect variation between samples. The DNA is quantified and equal amounts are aliquoted into tubes in eccess of the number of its targets in the samples, such as 10ug for a plasmid and 1ug for a PCR amplicon. These are denatured (NaOH and 95C) and added to the wells, were the vacuum sucks the water (with NaOH and NH3OAc) from underneath the membrane (nylon or nitrocellulose). When it is washed it can be dried and UV crossinked (faster) or baked. After that it can be prehybridized and then the sample in hybridization buffer (such as 50%formamide) can be added to the sacket at 48C to 60C.