Basic principles

Recombinant DNA technology provided the first ability to isolate gene sequences and prepare such molecules in unlimited abundance. The exquisite specificity of pairing between complementary strands of DNA and/or RNA in turn provided the means to measure the presence and quantity of specific genes and/or gene products in living cells and tissues. Perhaps the most common measurement performed in the analysis of gene expression is of the presence or abundance of specific mRNAs. Though this is now accomplished routinely, the major problem in the use of mononuclear phagocytes for determination of specific mRNA levels is presented by their unusually high content of ribonuclease activity, especially in cells which have been exposed to proinflammatory and/or activating stimuli. This problem may be compounded by a relatively low content of total RNA.

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