Protein Expression Prokaryotic cells C. crescentus JS1014

Get tips on using CelLytic™ B Cell Lysis Reagent to perform Protein isolation Bacteria - Synechococcus elongatus

Get tips on using CelLytic™ B Cell Lysis Reagent to perform Protein isolation Bacteria - Salmonella enterica

Get tips on using CelLytic™ B Cell Lysis Reagent to perform Protein isolation Bacteria - Pseudomonas aeruginosa

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Mouse heart

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Mouse aorta

Get tips on using Strep-Tactin Superflow Plus (10 ml) to perform Protein tag Purification of Strep-tagged proteins

Get tips on using Ni-NTA Fast Start Kit (6) to perform Protein tag Purification of His-tagged proteins

Get tips on using Penta·His Antibody, BSA-free (100 ug) to perform Protein tag Detection of His-tagged proteins

Get tips on using Tetra·His Antibody, BSA-free (100 µg) to perform Protein tag Detection of His-tagged proteins

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.