siRNA / RNAi /miRNA transfection Human Cells HESC

Get tips on using GeneJET Plasmid Miniprep Kit to perform Plasmid Isolation Proteus mirabilis

Get tips on using RNeasy 96 Kit (12) to perform mRNA / Ribonucleoprotein isolation / purification Ribonucleoprotein

Get tips on using Wizard® Plus Midipreps DNA Purification System Technical Bulletin to perform Plasmid Isolation Proteus mirabilis

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

Get tips on using ChargeSwitch™ Total RNA Cell Kit to perform RNA isolation / purification Bacteria - Gram negative Hemophilus influenzae

Get tips on using ChargeSwitch™ Total RNA Cell Kit to perform RNA isolation / purification Bacteria - Gram negative Haemophilus influenzae

Get tips on using QIAseq FX Single Cell RNA Library Kit (96) to perform Whole Transcriptome Amplification Virus

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.

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.