An essential pre-requisite to perform sound quantitative real-time polymerase chain reaction (qPCR) assays is to design outstanding primer pairs. This means they must have a good efficiency and be not prone to produce multiple amplicons or primer dimer products. To circumvent these issues, several softwares are available to help primer design. Although satisfactory computer-aided primer design ...

This paper describes a novel method of primer walking using octamer oligonucleotides to prime DNA sequencing reactions. Octamer sequencing is compatible with isotopic and fluorescent sequencing chemistry, reaction conditions are optimized such that the samples can be processed in parallel, and the procedure has the potential to be automated. This strategy is faster than the traditional primer w...

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 ...

Fingerprinting with molecular markers allows precise, objective and rapid variety identification. In this study, simple sequence repeat (SSR) markers were used to fingerprint the forty-eight main commercial rice cultivars under cultivation at present in Zhejiang Province, China, and identify indica and japonica subspecies including eight groups of closely related cultivars. Eighteen of the thir...

Using in vivo fidelity assays in which bacterial beta-galactosidase or green fluorescent protein genes served as reporters of mutations, we have identified a murine leukemia virus (MLV) RNase H mutant (Y586F) that exhibited an increase in the retroviral mutation rate approximately 5-fold in a single replication cycle. DNA-sequencing analysis indicated that the Y586F mutation increased the frequ...

The Polα/primase complex assembles the short RNA-DNA fragments for priming of lagging and leading strand DNA replication in eukaryotes. As such, the Polα polymerase subunit encounters two types of substrates during primer synthesis: an RNA:DNA helix and a DNA:DNA helix. The engagement of the polymerase subunit with the DNA:DNA helix has been suggested as the of basis for primer termination in e...

In search of a primer set that could be used to study Planctomycetes dynamics in lakes and especially via fingerprinting methods, e.g. denaturing gradient gel electrophoresis (DGGE), three existing specific primer sets, developed for marine and soil systems, have been tested on water samples from four freshwater ecosystems. The first primer set (PLA46F/PLA886R) allowed PCR amplification of Plan...

TG572 RFLP Probe: Five primers were designed around the RFLP probe on chromosome 7: P572F1, P572F2, P572F3, P572R1, and P572R2 (Table 1). All possible primer combinations produced a strong band with Heinz 1706 DNA between 600 bp and 1000 bp (Fig. 2). All of the primer combinations produced single bands with the exception of P572F2/P572R1, which produced two bands one at 400 bp and one at 900 bp...

target sequence is correct, the target sequence is amplified, and if the primer pair is in the wrong orientation relative to the amplified cDNA inserts, the target sequence is not amplified. This simple, linear pre-amplification of cDNAs in the library has the potential for wide use. This approach requires not only far fewer starting materials than conventional PCR cloning but also circumvents ...

Drosophila retrotransposons have been shown to have genes for enzymes similar to the reverse transcriptase of retroviruse. They may possibly be involved in genome replication on translocation (15). Identification was made of a primer tRNA for the putative reverse transcription of a Drosophila retrotransposon, 297, and its genomic counterparts using a homology to the putative primer binding site...