siRNA / miRNA gene silencing Human RMS

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.

Get tips on using Gibco™ DMEM, high glucose to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using BrainPhys™ Without Phenol Red to perform Stem cell Differentiation media Differentiation of Human iPSCs into Basal Forebrain cholinergic neurons (BFCN)

Get tips on using DuoSet® ELISA Development Systems to perform ELISA (kit) IL-6, IL-1β, IL-8 and TNF-α - -NA- Human -NA-

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human aortic smooth muscle cells (HOSMC)

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - BxPc-3 human primary pancreatic adenocarcinoma

Get tips on using CellTiter-Glo® Luminescent Cell Viability Assay to perform Cell cytotoxicity / Proliferation assay cell type - BxPc-3 human primary pancreatic adenocarcinoma

Get tips on using Cytoselect™ Cell Viability and Cytotoxicity Assay to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using TumorTACS™ In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells

Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells