siRNA / miRNA gene silencing Human Primary Human Aortic Endothelial Cells

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 PE Annexin V Apoptosis Detection Kit with 7-AAD to perform Apoptosis assay cell type - T-cells Mouse (OT-I)

Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM) with high glucose: Liquid to perform 3D Cell Culture Media U87MG cells- glioblastoma spheres

Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM)/ F12 1:1: Liquid to perform 3D Cell Culture Media U87MG cells- glioblastoma spheres

Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM) with low glucose: Liquid to perform Stem cell culture media hAdipose derived stem cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - Cancer cell lines Liver cancer cell lines Hepato cellular carcenoma (SMMC-7721, Huh7 & HepG2))

Get tips on using Jump In™ T-REx™ HEK 293 Kit to perform Protein expression and purification Mammalian cells - HEK 293 HER2

Get tips on using pMT/BiP/V5-His A, B, & C Drosophila Expression Vectors to perform Protein expression and purification Insect cells - S2 HER2

Get tips on using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® to perform RNA sequencing Mouse - ESCs (Embryonic Stem Cells)

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - BHK-21