Compuestos y reactivos bioquímicos

<p>The 25mM solution of MgCl2, (0.22μm membrane-filtered), is used for optimization of magnesium ion concentration in PCR.</p>

TRIzol-T Reagent is a ready-to-use reagent for the isolation of total RNA from cells and tissues. The reagent, a mono-phasic solution of phenol and guanidine isothiocyanate, is an improvement to the single-step RNA isolation method and gives high yields of Total RNA. During sample homogenization or lysis, TRIzol-T Reagent maintains the integrity of the RNA, while disrupting cells and dissolving cell components. Addition of chloroform followed by centrifugation, separates the solution into an aqueous phase and an organic phase. RNA remains exclusively in the aqueous phase. After transfer of the aqueous phase, the RNA is recovered by precipitation with isopropyl alcohol. After removal of the aqueous phase, the DNA and proteins in the sample can be recovered by sequential precipitation. Precipitation with ethanol yields DNA from the interphase, and an additional precipitation with isopropyl alcohol yields proteins from the organic phase. Copurification of the DNA may be useful for normalizing RNA yields from sample to sample.This technique performs well with small quantities of tissue (50-100 mg) and cells (5 × 106), and large quantities of tissue (≥1 g) and cells (>107), of human, animal, plant, or bacterial origin.

<p>10X PCR Buffer (with MgCl2 ) is recommended for all PCR applications with Taq DNA Polymerases (both recombinant and native ).</p>

SDS-PAGE is a method employed for separating proteins via electrophoresis, based on the principle that charged molecules migrate through a matrix in response to an applied electric field. The matrix utilized for this separation is polyacrylamide, which is derived from acrylamide, a compound that can be hazardous. Polyacrylamide is commonly used for the size-based separation of proteins and nucleic acids. The gel matrix forms through the free radical polymerization of acrylamide along with a crosslinking agent known as N, N-Methylenebisacrylamide. In this process, acrylamide monomers polymerize into long chains, initiated by a free radical-generating system, and these chains are linked by the cross-linker, resulting in a gel structure. This gel is crucial for effective electrophoretic separation, facilitating the analysis of biomolecules based on size.