siRNA / miRNA gene silencing Human hES cell line H1 (WA01)

Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - primary human coronary artery smooth muscle cells

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb to perform Cell cytotoxicity / Proliferation assay cell type - K562

Get tips on using 8 µm Chemotaxis Assays, 96-Well Format to perform Cell migration / Invasion cell type - MCF-10A

Get tips on using 8 µm Chemotaxis Assays, 24-Well Format to perform Cell migration / Invasion cell type - PANC-1

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - FADU

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 Xfect™ Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human pulmonary artery smooth muscle cells (HPASMC)

Get tips on using EpiQuik Dnmt3A Assay Kit to perform DNA methylation profiling Whole genome profiling - MCF-7, MDA-MB-453 human breast cancer

Get tips on using RPMI 1640 to perform Mammalian cell culture media HepG2

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - THP-1