Mu. Patwary et al., REVEALING GENETIC-MARKERS IN GELIDIUM-VAGUM (RHODOPHYTA) THROUGH THE RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) TECHNIQUE, Journal of phycology, 29(2), 1993, pp. 216-222
The recently developed random amplified polymorphic DNA technique was
evaluated as a method for characterizing isolates of the agarophyte Ge
lidium vagum Okamura. Reaction conditions for single primer polymerase
chain reaction were optimized to obtain a high degree of reproducibil
ity of the amplified bands generated from purified G. vagum DNA. A tot
al of 165 primers, including both (A + T)- and (G + C)-rich sequences,
was screened for DNA amplification using template DNA from a single G
elidium isolate. None of the 45 (A + T)-rich primers was positive (i.e
. band-producing). Of the (G + C)-rich primers, 47 were positive, gene
rating a total of 322 prominent amplification products for DNA from 13
different G. vagum isolates. Polymorphic DNA loci were detected by 37
of the primers. Unweighted pair-group arithmetic average cluster anal
ysis (UPGMA) of these loci was used to group the G. vagum isolates and
thereby determine which were most similar. G. latifolium, used as an
out-group for the UPGMA analysis, showed a high degree of dissimilarit
y.