TITLE

Nonradioactive Identification of Differential Display RT-PCR Products

AUTHOR(S)
Colmenares, Maribel; Gimenez, Carlos; Oropeza, Maira; De Gracia, Eva
PUB. DATE
March 2000
SOURCE
Plant Molecular Biology Reporter;2000, Vol. 18 Issue 1, p65
SOURCE TYPE
Academic Journal
DOC. TYPE
Abstract
ABSTRACT
Abstract. A simple, sensitive protocol for nonradioactive screening of DDRT-PCR candidates is described. Embryogenic and nonembryogenic cDNAs were prepared from mRNA isolated using paramagnetic particles. Reverse transcription was performed with biotinylated oligo-dT[sub (18)]. These labeled cDNAs were checked for integrity and used as a probe for screening DDRT-PCR candidates. Most of the DDRT-PCR candidates detected were from mRNA expressed at low levels, giving weak or undetectable signals, and stronger signals represented more highly expressed mRNAs. Our results demonstrate the usefulness of chemiluminescence detection to screen DDRT-PCR products. This strategy is an alternative method to radioactive detection procedures.
ACCESSION #
3871747

 

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