protein-expression-and-purification-bacteria-escherichia-coli-p19

Get tips on using pMT/BiP/V5-His A, B, & C Drosophila Expression Vectors to perform Protein expression and purification Insect cells - S2 HER2

Get tips on using BLOCK-iT™ Adenoviral RNAi Expression System, pAd/BLOCK-iT™-DEST RNAi Gateway Vector to perform shRNA gene silencing Mouse - P19 Foxm1

Get tips on using pMCY87 to perform Protein Expression Prokaryotic cells - E. coli PduP-E bacteriophage ϕX174

Get tips on using pMCY86 to perform Protein Expression Prokaryotic cells - E. coli PduP-E bacteriophage ϕX174

Get tips on using pMCY85 to perform Protein Expression Prokaryotic cells - E. coli PduP-E bacteriophage ϕX174

Get tips on using pET-32c to perform Protein Expression Prokaryotic cells - E. coli rpf-like protein

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.