DNA cloning is often used to select and amplify one DNA species from a mixture. However, the cloning process is complex and labor-intensive. We have developed a new two-step method for DNA sequencing directly from a mixture. The first is the introduction of a known oligonucleotide (common part) into the terminus of unknown DNA by ligation. The second is selective DNA sequencing using primers wi...

The random amplified polymorphic DNA method was used to distinguish strains of Xanthomonas campestris pv. pelargonii from 21 other Xanthomonas species and/or pathovars. Among the 42 arbitrarily chosen primers evaluated, 3 were found to reveal diagnostic polymorphisms when purified DNAs from compared strains were amplified by the PCR. The three primers revealed DNA amplification patterns which w...

background and objectives: dna ladder contains dna fragments of different length but with known size, used to determine the size of unknown dna molecules. different dna ladders are available for expected dna length. conserved sequences were selected for design of primers to generate dna fragments of known specific size. materials and methods: in this study, we describe a method by which dna lad...

OBJECTIVE The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. MATERIAL AND METHODS Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2...

We introduce quantitative polymerase chain reaction (qPCR) primers and multiplex end-point PCR primers modified by the addition of a single ortho-Twisted Intercalating Nucleic Acid (o-TINA) molecule at the 5'-end. In qPCR, the 5'-o-TINA modified primers allow for a qPCR efficiency of 100% at significantly stressed reaction conditions, increasing the robustness of qPCR assays compared to unmodif...

Cycle sequencing is the workhorse of DNA sequencing projects, allowing the production of large amounts of product from relatively little template. This cycling regime, which is aimed at linear growth of the desired products, can also produce artifacts by exponential amplification of minor side-products. These artifacts can interfere with sequence determination. In an attempt to allow linear but...

The ease and advantages of allelespecific PCR (AS-PCR) for SNP genotyping, along with the difficulty of assay specificity with certain mismatched DNA primers, have been described in prior studies (1–3). More recently, the use of real-time PCR detection formats has been described for one or both alleles of AS-PCR (4–6). These detection enhancement methods do not overcome the inherent difficultie...