Get tips on using pSilencer™ 4.1-CMV neo to perform shRNA gene silencing Human - SiHa AEG-1
Get tips on using SMARTpool: ON-TARGETplus TP63 siRNA to perform siRNA / miRNA gene silencing Human - A253 P36
Get tips on using CometAssay Electrophoresis System II to perform DNA Damage Assay Human Skin Fibroblast Cell (FSK)
Get tips on using CometChip Electrophoresis Starter Kit to perform DNA Damage Assay Human bronchial epithelial cells (hBE)
Get tips on using Comet SCGE assay kit to perform DNA Damage Assay Human bronchial epithelial cells (hBE)
Get tips on using SurePrint Human miRNA Microarrays to perform Microarray Human - Endometrial stromal cells miRNA-expression array (labelled)
Get tips on using JetPrime to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)
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.
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment