Recovery, purification, and cloning of high-molecular-weight DNA from soil microorganisms.

Soil Microorganisms of High-Molecular-Weight DNA from Recovery, Purification, and Cloning

Isolation of High Molecular Weight DNA from Forest Topsoil for Metagenomic Analysis

In this study, we employed a modified method to extract DNA from forest topsoil that was suitable for construction of large insert soil metagenomic library. The DNA extraction method used produced considerable DNA yield with DNA fragments ranging from 48 kb up to 290 kb. The recovery of soil DNA suitable for PCR and metagenomic library construction is difficult because soil DNA is often co-puri...

Rapid purification of HU protein from Halobacillus karajensis

The histone-like protein HU is the most-abundant DNA-binding protein in bacteria. The HU protein non-specifically binds and bends DNA as a hetero- or homodimer, and can participate in DNA supercoiling and DNA condensation. It also takes part in DNA functions such as replication, recombination, and repair. HU does not recognize any specific sequences but shows a certain degree of specificity to ...

Comparative Studies on the Extraction of Metagenomic DNA from Various Soil and Sediment Samples of Jammu and Kashmir Region in Prospect for Novel Biocatalysts

A modified and efficient method has been developed for extracting high molecular weight (HMW) DNA free of inhibitors (humic substances) from soil and sediment samples of Jammu and Kashmir region. The method includes precipitation of extracted soil and sediment DNA with 0.5 volumes of 50% polyethylene glycol (PEG) and 0.1 volumes of 1M NaCl (sodium chloride) followed by treatment with 2% CaCl2( ...

Cloning and Enhanced Expression of an Extracellular Alkaline Protease from a Soil Isolate of Bacillus clausii in Bacillus subtilis

in the detergent industry. In this study, the extracellular alkaline serine protease gene, aprE, from Bacillusclausii was amplified by PCR and further cloned and expressed in B. subtilis WB600 using the pWB980 expression vector. Protease activity of the recombinant B. subtilis WB600 harboring the plasmid pWB980/aprEreached up to 1020 U/ml, approximately 3-folds higher than the nativ...