shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y) Connexin 43

Get tips on using Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F12 to perform 3D Cell Culture Media PLC/PRF/5 cells-spheres

Get tips on using ApopTag® Fluorescein In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - Islets of langerhans (Beta cells)

Get tips on using Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham to perform Stem cell culture media NCH421K cells primary glioma

Get tips on using pwPICZalpha-DT390-bi-pIL-2-Gly to perform Protein Expression Eukaryotic cells - P. pastoris Porcine IL-2 fusion toxins

Get tips on using pFastBac-GP67-H6HA1-His-RhPV-IRES-EGFP to perform Protein Expression Eukaryotic cells - S. frugiperda HA1 of H6N1 AIV

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages

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.

Get tips on using MEBMTM Mammary Epithelial Cell Growth Basal Medium to perform 3D Cell Culture Media Mouse primary breast cancer ephitelial cells-Mammospheres

Get tips on using EBMTM-2 Endothelial Cell Growth Basal Medium-2 to perform Stem cell culture media Cord blood-derived endothelial cells(hCBiPS2)