Flotation as a tool for indirect DNA extraction from soil

The method proved to be an efficient alternative to direct extraction protocols where cell lysis is performed in the presence of soil particles. The environmental DNA extracted after flotation had high molecular weight and comparable yield as when using available commercial kits (3.5 mug DNA/g soil), and neither PCR nor restriction enzyme

Methods of enhancing the recovery of plasmid genes from

In this study we have investigated the use of flotation and filtration, singly and combined, to enhance the separation of plasmid containing liquors from neutralised lysates with very different levels of solids. Filtration of crude neutralised lysates, containing roughly 100 g l−1 solids, through various diatomaceous earth and cellulose precoat materials was invariably accompanied by severe

(PDF) Development of a Chemical-free Floatation

Oct 05, 2021 The quantitative eleme ntal ana lysis that has been per formed for both. ... at a recovery 80.1% can be achieved by cleaning the rougher concentrate five times in a conventional flotation cell

Subcellular Colocalization of the Cellular and Scrapie

neuroblastoma cells using the ice-cold Triton X-100 detergent method. (A) Distribution of PrP after lysis of N2a and ScN2a cells in ice-cold Triton X-100 and separation of CLDs by flotation into sucrose gradients. Aliquots of gradient fractions were subjected to immuno-blot analysis with the anti-PrP polyclonal R073 rabbit antiserum (26)

FLOTATION CHEMISTRY OF HEMATITE/OLEATE SYSTEM

Flotation A modified Hallimond cell with automatic control of flotation time and stirring intensity and time, was used for all flotation experiments. The flota-tion procedure consisted of desliming 0.8 gram of a sample till free of visible fines and transferring it to a cylinder to

Dimerization controls the lipid raft partitioning of uPAR

Nov 17, 2003 Cell lysis, flotation analysis, immunoprecipitation and immunoblotting For immunoblotting and co-immunoprecipitation experiments cells were lysed in RIPA buffer [0.1% sodium dodecyl sulfate (SDS), 1% Triton X-100, 1% deoxycholate, 0.15 M NaCl and 0.1 M Tris–HCl pH 7.6) containing a cocktail of protease inhibitors (Complete, Roche)

This item was submitted to Loughborough’s Institutional

13 flotation efficiency, and reduced the solids content in the plasmid liquor ten-fold. However, 14 strong reinforcement of the cell debris lattice by these microspheres hindered plasmid 15 release into the liquor beneath. 16 CONCLUSION: By incorporating magnetic or buoyant seeds during cell lysis we have

Cellular Protein fractionation for Mammalian Cells

Cell harvest and lysis 1) Collect cells and incubate in hypotonic lysis buffer (20mM Tris pH 7.5, 5mM MgCl2, 5mM CaCl2, 1mM DTT, 1mM EDTA, protease inhibitor). Incubate in lysis buffer for 30 minutes on ice. 2) Dounce homogenize the cells (40 strokes) and view cells under the microscope to verify that the membranes have been removed from the cells

Membrane Protein Extraction and Isolation | Thermo

89842. Reagent-based membrane protein extraction and membrane protein isolation. Reagent-based lysis has replaced traditional physical lysis as the method of choice for membrane disruption and extraction of membrane proteins. Reagent-based lysis methods do not require expensive, cumbersome equipment and protocols that are difficult to implement

Sludge flotation, its causes and control in granular

Jun 03, 2018 Sludge flotation is a commonly reported and long-standing issue hindering not only the widespread implementation of upflow anaerobic sludge bed (UASB)-type bioreactors in wastewater treatment but also the development of novel anaerobic/anoxic treatment processes such as anammox, partial denitrification, and biological sulfate reduction. This review attempts to address the instability of

Pre-oxidation enhanced cyanobacteria removal in drinking

The sequential introduction of KMnO 4 and Fe(II) can realize the moderate pre-oxidation and finally enhance the efficiency of coagulation and flocculation (Ma et al., 2014; Qi et al., 2020; Qi et al., 2016c). Fig. 2 shows the mechanisms of Mn(VII)-Fe(II) pre-treatment for algae removal by Al coagulation. The excessive inactivation of cyanobacterial cells can be prevented by the subsequent

(PDF) Investigating dissolved air flotation performance

Filaments of Plank- and taste and odour compounds by C/F/S, Himberg et al. tothrix rubescens and cells of M. aeruginosa were selected, the (1989), Lam et al. (1995) and Hrudey et al. (1999) reported the latter also as a surrogate of cyanobacterial colonies of prob- occurrence of cell lysis and subsequent release of such lematic size

US20110124101A1 - Methods and devices for producing

A scalable process and device for producing a bio molecule, in particular pharmaceutical grade plasmid DNA is described. The process includes the steps of alkaline lysis, neutralization and clarification and can be further extended. For separating the lysate and the precipitate an improved floatation method is disclosed. This method is based on attachment of CO 2 bubbles on the precipitate floe

Membrane Flotation Assay

The cell lysate is loaded on the bottom of the gradient, overlaid with the gradient, and membranes float up as the iodixanol gradient self-generates. The lipid content of membranes and the concentration of associated proteins will determine the separation of different membranes within the gradient

Combination of aqueous two-phase flotation and

Nov 08, 2021 A novel purification method (ATPF-ITC) which combining aqueous two-phase flotation (ATPF) with inverse transition cycling (ITC) was established and used to efficiently purify recombinant β-glucosidase (GLEGB) from cell lysis solution

Flotation as a tool for indirect DNA extraction from

Jun 09, 2010 The method proved to be an efficient alternative to direct extraction protocols where cell lysis is performed in the presence of soil particles. The environmental DNA extracted after flotation had high molecular weight and comparable yield as when using available commercial kits (3.5 μg DNA/g soil), and neither PCR nor restriction enzyme digestion of DNA were inhibited

Membrane Flotation Assay | Request PDF

The cell lysate is loaded on the bottom of the gradient, overlaid with the gradient, and membranes float up as the iodixanol gradient self-generates. The lipid content of membranes and the