A new method was developed to detect DNA methylation in the Citrus genome using random amplification coupled with restriction enzyme digestion. Genomic DNA from Citrus grandis (L.) Osb., Poncirus trifoliata (L.) Raf., and their F1 hybrid was amplified using 7 individual 10-mer random primers. Prior to amplification the DNA templates were digested with 2 pairs of restriction endonucleases (HpaII...

A chromosome walking method was improved in this work. The new method was named as nonspecific primer anchored PCR (NPA-PCR). Nested gene specific primers were designed based on the known region and long random primer using degeneracy oligonucleotides for nonspecific anchoring. Annealing temperatures were varied to control the priming. Target sequences were obtained by PCR with random primer an...

The infraspecific variability of the species Leishmania infantum is studied by using genetic markers generated by random amplified polymorphic DNA (RAPD). We have applied this technique, using 18 primers of arbitrary sequence, to 33 strains of the parasite belonging to 18 zymodemes isolated in different clinical forms and hosts. Other strains belonging to the species L. donovani, L. major, L. t...

ES cells. This phenomenon has been observed in our laboratory in a number of cases. Clones putatively identified as homologous recombinants by Southern analysis can be quickly checked for random insertion of extra copies of the targeting vector by PCR. Given the small difference in size between the two alleles and use of only two primers common to both WT and mutant DNA, the amplification rate ...

Random amplified polymorphic DNA (RAPD) is a simple, fast and cost effective method for assessing genetic diversity of plant varieties. The reproducibility of the RAPD amplification was affected by several factors, therefore causing misinterpretation. In this study, we analyzed the effect of changing the concentration of magnesium chloride, template genomic DNA and Taq DNA polymerase with the o...

Automated fluorescence-based DNA sequence analysis offers the possibility to undertake very large scale sequencing projects. Directed strategies, such as the specific-primer-directed sequencing approach ('gene walking'), should prove useful in such projects. Described herein is a study involving the use of this approach in conjunction with automated fluorescence detection on a commercial instru...

The genetic distance of 11 cotton genotypes varying in heat tolerance was studied using RAPD markers. Fifty-three random decamer primers were used for the estimation of genetic distance. Among the 53 RAPD primers, which were custom synthesized by GeneLink Inc., UK, 32 were polymorphic and 21 were monomorphic. The 32 polymorphic primers produced 273 fragments, with a mean of 8.3 fragments per pr...

Fusarium verticillioides (F. verticillioides) is not only a primary pathogen of maize, but also can causedisease in other crops such as sorghum. Pathogenicity is related to mycotoxin production such as fumonisin.In the present study, 24 isolates of F. verticillioides, which were previously identified by phenotype basedmethods, were re-identified using restriction fragment length polymorphism (R...

A previous study reported the cloning of a putative Prevotella nigrescens-specific DNA probe, Pn23, using random shotgun method. The present study evaluated the species-specificity of Pn23 for P. nigrescens using the clinical strains of Prevotella intermedia and P. nigrescens to develop Prevotella nigrescens-specific polymerase chain reaction (PCR) primers. The specificity was tested against 10...

For the first time, genetic variability and differentiation among Ten Synconium white butterfly populations originating from tissue cultured plants maintained in pro-trays under the poly house & green house conditions in Indo American hybrid Seed Company, Bangalore were examined. Random Amplified Polymorphic DNA (RAPD) marker data were obtained and analysed with respect to genetic diversity. Th...