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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.4/1.2


TITLE: PCR ANALYSIS FOR HTLV 1,2


HTLV-1 and HTLV-2 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 pol region of HLTV-1/-2 (SK 110 and SK 111) (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. Kwok S, Kellogg D, Ehrlich G, Poiesz B, Bhagavati S, Sninsky JJ: Characterization of a sequence of human T cell leukemia virus type I from a patient with chronic progressive myelopathy. J Infect Dis 158: 1193-1197 (1988).


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