rna-isolation-purification-cells-immortalized-bxpc-3

Get tips on using ChargeSwitch™ Total RNA Cell Kit to perform RNA isolation / purification Bacteria - Gram negative Haemophilus influenzae

Get tips on using NucleoSpin® RNA Blood to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

Get tips on using 14-3-3ζ siRNA(h) to perform siRNA / miRNA gene silencing Human - Caco-2 14‐3‐3ζ

Get tips on using E.Z.N.A.® Total RNA Kit I to perform RNA isolation / purification Bacteria - Gram positive Staphylococcus epidermidis

Get tips on using Direct-zol™ RNA MiniPrep Plus to perform RNA isolation / purification Bacteria - Gram negative Borrelia burgdorferi

Get tips on using Direct-zol™ RNA MiniPrep Plus to perform RNA isolation / purification Bacteria - Gram negative Escherichia coli

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray RNA amplification & Labeling - Mouse brain tissue Biotin

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.