Development of yeast artificial chromosome (Y AC) vectors, molecular cloning of large segments of chromosomal DNA, and their propagation in yeast cells has become feasible. Overlapping Y AC provides a route to the development of physical maps of entire mammalian chromosomes. A rapid method was developed to isolate and sequence termini of Y AC inserts quickly. The Y AC clone is digested wit...

Salmonella enterica subspecies enterica serovar Gallinarum biovar Gallinarum is the causative agent of fowl typhoid in chickens, outbreaks of which have devastated poultry populations in Korea since 1992. In order to identify genetic differences among S. Gallinarum isolates, bacteria were examined using the random amplified polymorphic DNA (RAPD) method. Of 13 arbitrary primers screened initial...

We extended the concept of fluorescent microsatellite genotyping with a single-universal tailed primer to the simultaneous use of three different tailed primers to allow multiplexed 4-color detection for medium throughput genotyping of plant species. The method was tested on Eucalyptus DNA samples using three forward primer sequences of human microsatellite markers labeled with different fluore...

Hydrocarbonoclastic bacteria (HCB) play a key role in the biodegradation of oil hydrocarbons in marine and other environments. A small number of taxa have been identified as obligate HCB, notably the Gammaproteobacterial genera Alcanivorax, Cycloclasticus, Marinobacter, Neptumonas, Oleiphilus, Oleispira, and Thalassolituus, as well as the Alphaproteobacterial genus Thalassospira. Detection of H...

Rolling circle amplification (RCA) catalyzed by φ29 DNA polymerase offers a simple method for DNA amplification in the presence of a circular DNA template and its complimentary primer. RCA continuously produces long single-strand DNA using the strand displacement activity of polymerase during DNA synthesis. This property allows one to monitor the progress of a reaction by means of electrophores...

Total RNA from wheat spikes infected with F. graminearum Fg2 was extracted and the mRNA was purified. Switching Mechanism at 5' end of the RNA Transcript (SMART) technique and CDS Ill/3' primer were used for first-strand cDNA synthesis using reverse transcriptase by RT-PCR. Primer extension polymerase chain reaction was used to construct the double-strand cDNA that was digested by proteinase K,...

A rapid and efficient procedure for site specific mutagenesis is described. A double primed synthesis with a 17-mer mismatch primer and a "universal" 15-mer M13 sequencing primer was used to introduce a T to A transversion into an ompF signal peptide gene cloned in the M13mp8 vector. The two primers were annealed to the circular single stranded M13 template. After a short extension and ligation...

In the present study, we report the occurrence of cry8 positive isolates of Bacillus thuringiensis (Bt) in selected white grub, Holotrichia serrata F. (Coleoptera: Scarabaeidae), endemic soils of sugarcane ecosystem and other places in Tamil Nadu. Out of the 66 soil samples collected and screened for white grub specific Bt, 74 isolates of the bacterium, all containing only spherical crystal tox...

Current methods of understanding microbiome composition and structure rely on accurately estimating the number of distinct species and their relative abundance. Most of these methods require an efficient PCR whose forward and reverse primers bind well to the same, large number of identifiable species, and produce amplicons that are unique. It is therefore not surprising that currently used univ...

This study evaluated the effects of three metal primers and one multi-mode adhesive system on the shear bond strength (SBS) of a flowable composite resin to nickel-chrome metal alloy (Ni-Cr). Ninety plates were cast from Ni-Cr and divided in nine groups (n=10). The surfaces were sandblasted with Al2O3 and primed with three adhesive primers: Alloy Primer (AP), Universal Primer (TP) and RelyX Cer...