The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.
Get tips on using RNAqueous™ Total RNA Isolation Kit to perform RNA isolation / purification Tissue - Mouse Uterus
ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - Rat Blood / Serum / Plasma / Buffy coat
Get tips on using RNeasy Lipid Tissue Mini Kit to perform RNA isolation / purification Tissue - mouse kidney tissue
Get tips on using RNeasy Lipid Tissue Mini Kit to perform RNA isolation / purification Tissue - human adipose tissue
When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.
Get tips on using PureLink™ FFPE RNA Isolation Kit to perform RNA isolation / purification Tissue - Human Kidney
Get tips on using High Pure FFPET RNA Isolation Kit to perform RNA isolation / purification Tissue - Human Kidney
Get tips on using High Pure FFPET RNA Isolation Kit to perform RNA isolation / purification Tissue - Human Breast
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment