The efficacy of random primer-pair arrays compared to conventional RAPD method with a single decamer primer was evaluated using DNA from two species of Cucumis. The banding patterns of amplicons revealed enhanced utility of primer-pair arrays over conventional RAPDs, producing more bands and a higher degree of polymorphism, both at intra- and inter-specific levels. Amplification produced by bot...

Due to lack of morphological methods to identify sex at early stage in the plants with long juvenile period the application of molecular markers is expected to facilitate breeding program. The objective of this study is to identify molecular markers linked to sex determination of the plant Simarouba glauca which assists in crop improvement program. Random amplified polymorphic DNA primers were ...

Molecular genetics, and in particular the development of the polymerase chain reaction (PCR), has provided a way to distinguish these organisms. One primer pair specific for the genus Giardia is available (Mahbubani et al. 1991) as is a primer pair specific for G.intestinalis, (Mahbubani et al. 1992). During experiments in this laboratory, designed to compare strains and species of Giardia usin...

BACKGROUND Polymerase chain reaction (PCR) is one of the most important developments in modern biotechnology. However, PCR is known to introduce biases, especially during multiplex reactions. Recent studies have implicated the DNA polymerase as the primary source of bias, particularly initiation of polymerization on the template strand. In our study, amplification from a synthetic library conta...

Microsatellites or simple sequence repeats provide an attractive source of genetic polymorphisms for both mammals and plants (1). Microsatellites can be detected by PCR using minute amounts of starting material and they provide codominant markers with a high degree of allelic polymorphism. However, these markers are obtained through a difficult and labor-intensive procedure. In contrast, RAPD m...

INTRODUCTION In this method, an oligodeoxynucleotide primer hybridized to mRNA is extended by an RNA-dependent DNA polymerase to create a cDNA copy that can be amplified by PCR. Depending on the purpose of the experiment, the primer for first-strand cDNA synthesis can be specifically designed to hybridize to a particular target gene, or a general primer such as oligo(dT) can be used to prime cD...

Fusarium verticillioides is a pathogen of agriculturally important crops, especially maize. It is considered one of the most important pathogens responsible for fumonisin contamination of food products, which causes severe, chronic, and acute intoxication in humans and animals. Moreover, it is recognized as a cause of localized infections in immunocompetent patients and disseminated infections ...

We characterized single primer amplification reaction (SPAR) molecular markers from 20 genotypes of Anthurium andraeanum Lind., including 3 from commercial varieties and 17 from 2 communities in the State of Espírito Santo, Brazil. Twenty-four SPAR, consisting of 7 random amplified polymorphic DNA and 17 inter-simple sequence repeat markers were used to estimate the genetic diversity of 20 Anth...

We have developed a novel method to clone and sequence minute quantities of DNA. The method was applied to sequence a 180 kb plasmid pNL1. The first step was the production of a size distributed population of DNA molecules that were derived from the 180 kb plasmid pNL1. The first step was accomplished by a random synthesis reaction using Klenow fragment and random hexamers tagged with a T7 prim...

Here we report a new methodology to study trace amounts of DNA of unknown sequence using a two-step PCR strategy to amplify and clone target DNA. The first PCR is carried out with a partial random primer comprised of a specific 21-nucleotide 5' sequence, a random heptamer, and a 3' TGGC clamp. The second PCR is carried out with a single 19-nucleotide primer that matches the specific 5' sequence...