Protein refolding procedure generally begins with aggregated protein extraction anu dissolving in high concentration chaotropic
agents such as urea and guanidine hydrochloride (GuHCl).
Many DNA extraction protocols depend on silica-based adsorbents to selectively bind and elute DNA in the presence of chaotropic
agents and low ionic-strength buffers respectively.
It is known that urea is a chaotropic
agent, efficient in the rupture of hydrogen bonds, denaturing proteins by breaking the noncovalent and ionic links between aminoacid residues .
(23-25) In this case, by utilizing chaotropic
conditions generated within the collected sample, human cells are selectively lysed, after which, enzyme-mediated digestion of human DNA and extracellular pathogen DNA occurs, followed by sedimentation of intact pathogen cells with the aid of centrifugation.
The methodology of phenol-chloroform extraction and DNA purification utilized the Brazol, which has in its composition in addition to phenol, guanidine thiocyanate, a chaotropic
agent that inactivates endonucleases and prevents DNA binding to other molecules and facilitates the separation of cellular debris [23-26].
Lytic enzymes, chaotropic
agents, and different types of detergents are the main components of chemical lysis, while mechanical method disrupts the cells by grinding, shearing, bead beating, and shocking .
Briefly, specimens were digested under denaturing conditions at elevated temperatures and then lysed in the presence of chaotropic
In the following step, the lysis fluid containing DNA/RNA is transferred to a chaotropic
buffer containing fresh silica beads.
In brief, the sputum samples were mixed with 2-3 volumes of the USP solution, which contained the following chemicals: 4-6 M guanidinum hydrochloride (a chaotropic
agent which disrupts the hydrogen bonds), 50 mM Tris chloride (pH 7.5), 25 Mm Ethylene Diamine Tetra Acetic acid (EDTA), 0.5% sarkosyl, and 0.1-0.2 M Beta mercaptoethanol These ingredients together brought about mucolysis and acted as detergents.
Protein amounts (based on the Bradford assay) were first dried by vacuum concentration and then resuspended in a strong chaotropic
solution made of the following: 7 M urea, 2 M thiourea, 5% (w/v) SDS, 40 mM Tris-HCl, pH 6.8,0.5 mM ethylenediaminetetraacetic acid (EDTA), 20% (v/v) glycerol, and 50 mM dithiothreitol (DTT) in a ratio of 0.25 [micro]g of protein per [micro]L of solution .
acids such as guanidine hydrochloride or guanidinium thiocyanate were discovered to protect NA from nucleases because of their potent protein denaturing properties (16).