Methods of extracting, amplifying and detecting a nucleic acid f

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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424520, 435803, 436177, 935 77, 935 78, C12Q 168, G01N 110

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053344995

ABSTRACT:
A nucleic acid is rapidly extracted from whole blood or a peripheral blood mononuclear cell (PBMC) fraction thereof. Extraction from the PBMC fraction is accomplished by heating the fraction at or near the boiling point of water for a few minutes and recovering the extracted nucleic acid. This rapid method is particularly useful for extracting DNA for the detection of genetic diseases or infectious agents, such as HIV-I. Whole blood can likewise be heated after it is mixed with a salt solution containing a polysaccharide, such as dextran. The extracted nucleic acid is then recovered from the heated mixture. Nucleic acids extracted in this way are available for amplification using a polymerase chain reaction. Where the presence of a specific gene is to be determined for diagnostic purposes, it can be extracted as described above and subjected to suitable amplification and detection steps.

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