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A microassay for detection of DNA and RNA in small numbers of plant cells
Authors:Anne Crossway  Catherine M. Houck
Affiliation:(1) Calgene, Inc., 1920 Fifth St., 95616 Davis, CA, U.S.A.
Abstract:Summary A microtechnique for the detection of DNA or RNA in small numbers of plant cells (1–50) has been developed using cauliflower mosaic virus (CaMV) infection of turnip as a model system. Both DNA and RNA extracted from 10 mesophyll protoplasts from CaMV-infected plants can be detected by hybridization using a radioactive probe made from cloned CaMV DNA (pCaMV10). No hybridization above background was detected in extracts of protoplasts from uninfected plants. At least 0.15 pg (11 000 molecules) of purified pCaMV10 DNA can be detected. This method is superior to existing lsquomacrorsquo techniques for nucleic acid detection as smaller amounts of tissue are required and the detection is approximately 100-fold more sensitive. re]19850326 rv]19850530 ac]19850611
Keywords:cauliflower mosaic virus  microtechnique  nucleic acid hybridization
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