RNA isolation / purification Tissue Human

Get tips on using QIAamp 96 DNA QIAcube HT Kit (5) to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using QIAamp Fast DNA Stool Mini Kit (50) to perform DNA isolation / purification Tissue - fecal sample

Get tips on using MaXtract High Density (100 x 15 ml) to perform DNA isolation / purification Tissue - small intestine

Get tips on using AllPrep PowerViral DNA/RNA Kit (50) to perform DNA isolation / purification Viral

Get tips on using PureLink™ RNA Mini Kit to perform RNA isolation / purification Cells - primary canine coronary artery smooth muscle cells

Get tips on using E.Z.N.A.® Total RNA Kit I to perform RNA isolation / purification Cells - primary bovine umbilical vein endothelial cells

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Get tips on using VWR Life Science RiboZol™ RNA Extraction Reagent to perform RNA isolation / purification Cells - primary rat brain microvascular endothelial cells