siRNA / miRNA gene silencing Human 501 Mel and SK Mel 28 FANCD2

Get tips on using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Human - Deletion K562 c-Myb gene

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 Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Immortalized cell lines iPSC

Get tips on using Proteome Profiler™ Human Apoptosis Array Kit to perform Apoptosis assay cell type - Array of apoptotic proteins

Get tips on using Human RBP4/Retinol Binding Protein 4 PicoKine™ ELISA Kit to perform ELISA Human - RBP4

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Primary cells CD8+ T cells

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Primary cells CD4+ T cells

Get tips on using Monoclonal Mouse Anti-Human Ki-67 Antigen (Dako Omnis) Clone MIB-1 to perform Immunohistochemistry Human - Ki-67