Protein Expression Prokaryotic cells R. erythropolis

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 CelLytic™ B Cell Lysis Reagent to perform Protein isolation Bacteria - Bacillus cellulosilyticus

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using Proteome Profiler™ Human Apoptosis Array Kit to perform Apoptosis assay cell type - Array of apoptotic proteins

Get tips on using QIAexpress Type IV Kit to perform Protein tag Purification of His-tagged proteins

Get tips on using Penta·His HRP Conjugate Kit to perform Protein tag Detection of His-tagged proteins

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

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