siRNA / RNAi /miRNA transfection Human Cells HESC

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary rat lymph node cells

Get tips on using DMEM/F-12, no phenol red to perform 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using ROS-ID® Total ROS detection kit to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - Human osteosarcoma cancer cells

Get tips on using FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform Apoptosis assay cell type - Human endometrial stromal cells

Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - HEK293 human embryonic kidney cells

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 CellROX™ Deep Red Reagent, for oxidative stress detection to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

Get tips on using Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to perform Live / Dead assay yeast - Clostridium sporogenes