as a template for PCR. Sequencing of the plasmid DNA can now be performed using two PCR primers flanking the MCS of the vector (we used M13 universal forward primer and M13 universal reverse primer; Figure 1). If the unique primer is set 100–150 bp from the end of the known sequence, sufficient overlapping sequence is generated to align the sequences unambiguously. This method was used in our l...

Species diversity of metazoan bulk samples can be rapidly assessed using Cytochrome c oxidase I (COI) metabarcoding. However, in cases where only degraded DNA is available, e.g. from poorly conserved museum specimens, eDNA filtered from water or gut content analyses, universal primer sets that amplify only a short COI fragment are advantageous. Using PrimerMiner, we optimised two universal prim...

OBJECTIVES To evaluate the bonding of resin-cement to yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) via silica coating followed by silanization, and three one-bottle universal adhesives, with or without prior conditioning using a zirconia primer. METHODS Y-TZP specimens (n=160) were conditioned by tribochemical silica coating and silanization (CS), or alumina sandblasting with one...

Free-living nitrogen-fixing prokaryotes (diazotrophs) are ubiquitous in soil and are phylogenetically and physiologically highly diverse. Molecular methods based on universal PCR detection of the nifH marker gene have been successfully applied to describe diazotroph populations in the environment. However, the use of highly degenerate primers and low-stringency amplification conditions render t...

Genetic variability and relationships among ten rice varieties were studied by using 20 decamer random primers and SDSPAGE methods. Out of 22, 20 Random Amplified Polymorphic DNA (RAPD) primers revealed polymorphism, while the remaining 2 primers showed no reaction. The primers produced a total of 109 bands, of which 62 (56.88%) were polymorphic. The number of polymorphic fragments for each pri...

SSR or microsatellite markers are proved to be ideal for making genetic maps, assisting selection and studying genetic diversity in germplasm. SSR markers are playing important role to identify gene for salt tolerance that can be helpful for plant breeders to develop new cultivars. The experiment was conducted during the period from July 2009 to November 2010 in the experimental field and Biote...

Our understanding of thermophile diversity is based predominantly on PCR studies of community DNA. "Universal" and domain-specific rRNA gene PCR primers have historically been used for the assessment of microbial diversity without adequate regard to the degree of specificity of primer pairs to different prokaryotic groups. In a reassessment of the published primers commonly used for "universal"...

The objective of the present study is to detect genetic impurity in the seed lots of CMS lines, restorers and hybrids and to identify signature markers to differentiate parents and hybrids through DNA-based assays. Furthermore, attempts have been made to find out an alternative to Grow-Out-Test, which is very tedious, time consuming and used conventionally for seed genetic purity testing since ...

New bioactive proteins need to be screened from various microorganisms for the increasing need for industrial and pharmaceutical peptide, proteins, or enzymes. A novel polymerase chain reaction (PCR) method, restriction site-dependent PCR (RSD-PCR), was designed for rapid new genes cloning from genomic DNA. RSD-PCR strategy is based on these principles: (i) restriction sites disperse throughout...