DNA isolation / purification Plants

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized A549

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized HeLa

Get tips on using Nucleic Acid Purification to perform Plasmid Isolation Lactococcus lactis

Get tips on using Nucleic Acid Purification to perform Plasmid Isolation DH10Bac (Bacmid)

Get tips on using Gentra Puregene Cell Kit Plus (6.7 x 109) to perform DNA isolation / purification Cells - Immortalized cell lines H1 hESc

Get tips on using Absolutely Total RNA Purification Kits to perform RNA isolation / purification Cells - primary rat cardiac fibroblasts

Get tips on using Total RNA Purification Plus Kit to perform RNA isolation / purification Bacteria - Gram negative Vibro cholerae

Get tips on using XcelGen Fungal RNA Isolation Mini Kit to perform RNA isolation / purification Fungal

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.

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.