Get tips on using PowerPlex® 18D System to perform Cell line authentication Human iPSC cells derived from peripheral blood mononuclear cells
Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines Chang Liver cells
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized HeLa, CaSki and C33A (Cervical cancer cells)
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 PureLink™ RNA Mini Kit to perform RNA isolation / purification Cells - primary canine coronary artery smooth muscle cells
Get tips on using E.Z.N.A.® Total RNA Kit I to perform RNA isolation / purification Cells - primary bovine umbilical vein endothelial cells
Get tips on using Zombie Aqua™ Fixable Viability Kit to perform Live / Dead assay mammalian cells - human peripheral blood mononuclear cells
Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.
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 Live/Dead Cell Double Staining Kit to perform Live / Dead assay mammalian cells - MCF-7 human breast cancer cells
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