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 CM-H2DCFDA (General Oxidative Stress Indicator) to perform ROS assay cell type - CHO-K1
Get tips on using CM-H2DCFDA (General Oxidative Stress Indicator) to perform ROS assay cell type - 3T3-L1
Get tips on using GeneJET RNA Purification Kit to perform RNA isolation / purification Cells - primary human renal artery smooth muscle cells
Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - C2C12 myogenin
Get tips on using GenElute™ Mammalian Total RNA Miniprep Kit to perform RNA isolation / purification Cells - immortalized A2780
Get tips on using GenElute™ Mammalian Total RNA Miniprep Kit to perform RNA isolation / purification Cells - immortalized HeLa
Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - Neuroblastoma 2a Epac1
Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Human - Insertion IMR-32 SULT4A
Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Dog - Point mutation MDCK SCRIB
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment