Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency.

Bacterial competent cells are essential for cloning, construction of DNA libraries, and mutagenesis in every molecular biology laboratory. Among various transformation methods, electroporation is found to own the best transformation efficiency. Previous electroporation methods are based on washing and electroporating the bacterial cells in ice-cold condition that make them fragile and prone to death. Here we present simple temperature shift based methods that improve DNA transformation and recombineering efficiency in E. coli and several other gram-negative bacteria thereby economizing time and cost. Increased transformation efficiency of large DNA molecules is a significant advantage that might facilitate the cloning of large fragments from genomic DNA preparations and metagenomics samples

J Agric Food Chem

Pulsed electric field (PEF) treatment is an emerging technology that is arousing increasing interest in vinification processes for its ability to enhance polyphenol extraction performance. The aim of this study was to investigate the effects of PEF treatment on grape skin histocytological structures and on the organization of skin cell wall polysaccharides and tannins, which, until now, have been little investigated. This study relates to the effects of two PEF treatments on harvested Cabernet Sauvignon berries: PEF1 (medium strength (4 kV/cm); short duration (1 ms)) and PEF2 (low intensity (0.7 kV/cm); longer duration (200 ms)). Histocytological observations and the study of levels of polysaccharidic fractions and total amounts of tannins allowed differentiation between the two treatments. Whereas PEF1 had little effect on the polyphenol structure and pectic fraction, PEF2 profoundly modified the organization of skin cell walls. Depending on the PEF parameters, cell wall structure was differently affected, providing variable performance in terms of polyphenol extraction and wine quality