siRNA / miRNA gene silencing Human Calu-3

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.

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

Get tips on using High Pure miRNA Isolation Kit to perform RNA isolation / purification Tissue - Human Bone marrow

Get tips on using Anti-PI 3 Kinase p85 alpha (phospho Y607) antibody to perform Autophagy assay cell type - HepG2

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Mouth

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Larynx

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Eye

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Esophagus

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human osteoblasts

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Salivary glands