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Category Archives: Biology

biology

Western Blotting Hybridization Protocol

Sample protein preparation Monolayer Cells Grow cells to 80-90% confluency in a 100 mm dish, remove culture medium and rinse cell monolayer with 1x PBS at room temperature. Add 0.3-0.5 ml lysis buffer enriched with protease inhibitor cocktail to the monolayer cells in the plate. Scrape the cells off and transfer the lysate to an…

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In situ hybridization

A modern approach to the specific location of genes on chromosomes is a technique for the hybridization of DNA and RNA “in situ.” With this procedure, specific radioactive RNA or DNA (known as probes) can be isolated (or synthesized “in vitro”) and then annealed to chromosomes which have been treated in such a manner that…

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Working with DNA

DNA is a big molecule, but it’s still so small that there is no way to physically manipulate DNA. It is all done chemically using enzymes like reverse transcriptase, polymerase chain reaction (PCR), sequence analysis, and restriction enzymes. Reverse transcriptase makes DNA copies of RNA, PCR makes multiple copies of a particular gene and restriction enzymes cut…

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