Guidelines
CELL LINES
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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES
REFERENCE NO: AHC/1998/3/3.4/1.1
TITLE: PCR ANALYSIS FOR HIV
HIV DNA sequences integrated into the cellular genome (proviruses) are detected using the PCR amplification. DNAs from cell lines are extracted using proteinase K digestion followed by standard organic extractions. Specific oligonucleotide pairs from the gag region of HIV-1 (SK 38 and SK 39) (1) are used in a hot start PCR reaction. After 30 cycles at 95 C for 1 min, 55 C for 1 min and 72 C for 1 min the amplified products are identified by agarose gel electrophoresis and visualized by ethidium bromide intercalation.
References:
1. Kellogg D, Kwok S: Detection of human immunodeficiency virus. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds. PCR Protocols. A Guide to Methods and Applications. Academic Press, San Diego, 1990, pp. 337-347.
Guidelines prepared for CABRI by CERDIC, DSMZ, ECACC, INRC, November 1998
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