DNA

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 HT 8-oxo-dG ELISA Kit II to perform DNA Damage Assay MIA PaCa-2

Get tips on using OxiSelect™ Comet Assay Kit (3-Well Slides) to perform DNA Damage Assay MCF 10A

Get tips on using OxiSelect™ Comet Assay Kit (3-Well Slides) to perform DNA Damage Assay Hep G2

Get tips on using OxiSelect™ Comet Assay Kit (3-Well Slides) to perform DNA Damage Assay HEK 293T

Get tips on using JetPrime to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)

Get tips on using PolyMag Neo to perform DNA transfection Mammalian cells - Immortalized cell lines SH-SY5Y

Get tips on using EpiTect MSP Kit to perform DNA methylation profiling Gene specific profiling - HeLa MEG3

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - HepG2 Fibrinogen

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - SKOV3 AGR2