siRNA / miRNA gene silencing Rat H19-7

Get tips on using Subcellular Protein Fractionation Kit for Cultured Cells to perform Protein isolation Mammalian cells - Rat_Circumvallate papillae

Get tips on using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents to perform Protein isolation Mammalian cells - Rat_Liver

Get tips on using GRP 78 CRISPR Knockout and Activation to perform CRISPR Human - Activation GRP 78

Get tips on using Alexa Fluor 700-labeled anti-CD16 to perform Flowcytometry CD16 - Mouse /IgG1, kappa Human Alexa Fluor 700

Get tips on using Anti-Human CD56 (NCAM) APC-eFluor® 780 to perform Flowcytometry CD56 (NCAM) - Mouse / IgG1, kappa Human APC-eFluor 780

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 ATP5A1 Monoclonal Antibody (7H10BD4F9) to perform Western blotting ATP5A

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb to perform Autophagy assay cell type - RAW 264.7

Get tips on using Bax Antibody (4H32): sc-70407 to perform Western blotting Bax

Get tips on using Anti-Fas Antibody, clone 7C10 to perform Western blotting Fas