Applied Biosystems

About: Applied Biosystems is a based out in . It is known for research contribution in the topics: Mass spectrometry & Nucleic acid. The organization has 1521 authors who have published 1579 publications receiving 285423 citations.

Recombinant reverse transcriptases

Abstract: The present invention relates to a gene that encodes a hyperactive reverse transcriptase having DNA polymerase activity and substantially reduced RNase H activity, vectors containing the gene and host cells transformed with the invention. The present invention also includes a method of producing the hyperactive reverse transcriptase, producing cDNA from mRNA using the reverse transcriptase of the invention, kits and assay templates made using the hyperactive reverse transcriptase.

Biochemical characterization of DNA-binding proteins from Pyrobaculum aerophilum and Aeropyrum pernix

Abstract: Several representatives of the Crenarchaeal branch of the Archaea contain highly abundant, small, positively charged proteins exemplified by the Sso7d protein from Sulfolobus solfataricus. These proteins bind to DNA in a non-sequence-specific manner. Using publicly available genomic sequence information, we identified a second class of small Crenarchaeal DNA-binding proteins represented by the Pyrobaculum aerophilum open reading frame 3192-encoded (Pae3192) protein and its paralogs. We investigated the biochemical properties of the Pae3192 protein and an orthologous protein (Ape1322b) from Aeropyrum pernix in side-by-side experiments with the Sso7d protein. We demonstrate that the recombinant Ape1322b, Pae3192 and Sso7d proteins bind to DNA and that the DNA-protein complexes formed are slightly different for each protein. We show that like Sso7d, Pae3192 constrains negative supercoils in DNA. In addition, we show that all three proteins raise the melting temperature of duplex DNA upon binding. Finally, we present the equilibrium affinity constants and kinetic association constants of each protein for single-stranded and double-stranded DNA.

Single sheet seal applicator and cartridge

Abstract: An automated seal applicator for applying a sealing cover to a microplate. The microplate can comprise a plurality of wells for receiving an assay. The automated seal applicator can comprise a housing and a sealing cover cartridge containing a sealing cover releasably carried on a carrier liner. The sealing cover can be at least partially contained within the sealing cover cartridge. A sealing cover drive system can engage the sealing cover and/or the carrier liner and maintain a predetermined alignment of the sealing cover relative to the microplate.

Lifetimes of mRNAs for clock-regulated proteins in a dinoflagellate.

Abstract: Both pulsed and continuous applications of the RNA polymerase II inhibitor thiolutin cause a dramatic but reversible loss of bioluminescence and its overt rhythmicity in cells of the dinoflagellate Lingulodinium polyedrum (formerly Gonyaulax polyedra). Such cells remain alive, and the rhythm resumes after an interval, the length of which depends on the concentration of thiolutin used. The period and phase of the resumed rhythm were not systematically altered following such treatments, and the effects were not different at different circadian phases. For three different genes, luciferin binding protein (lbp), luciferase (lcf), and glyceraldehyde‐3‐phosphate dehydrogenase (gapdh), which are circadian‐regulated at the level of translation, the amounts of their mRNAs were determined by Northern blots for times up to 12.5h following the addition of 1.5 µM thiolutin. Consistent with previous reports that their abundances do not change with circadian time, their levels remained high for several hours after thiol...

5 – Quantitation of Nucleic Acids

Abstract: This chapter focuses on quantitation of nucleic acids by applying various methods. Any experiment requiting manipulation of a nucleic acid most likely also requires its accurate quantitation to ensure optimal and reproducible results. The nitrogenous bases positioned along a nucleic acid strand absorb ultraviolet (UV) light at a wavelength of 260 nm (nanometers). At this wavelength, light absorption is proportional to nucleic acid concentration. This relationship is so well characterized that UV absorption is used to accurately determine the concentration of nucleic acids in solution. Quantitation is performed by taking absorbance measurements at 260 nm, 280 nm, and 320 nm. Absorbance at 260 nm is used to specifically detect the nucleic acid component of a solution. The chapter also describes oligonucleotide quantitation. Many laboratories express an amount of oligonucleotides in terms of optical density (OD) units.