Get tips on using Cell Counting Kit-8 to perform Live / Dead assay mammalian cells - rat nucleus pulposus
Get tips on using Cellstain-Double Staining Kit to perform Live / Dead assay mammalian cells - HepaRG human hepatoma
Get tips on using Cell Counting Kit-8 (CCK-8) to perform Live / Dead assay mammalian cells - MMQ
Get tips on using Zombie Aqua™ Fixable Viability Kit to perform Live / Dead assay bacteria - Staphylococcus aureus
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 JetPrime to perform DNA transfection Mammalian cells - Immortalized cell lines PANC-1
Get tips on using NeuroMag to perform DNA transfection Mammalian cells - Immortalized cell lines SH-SY5Y
Get tips on using EvaGreen® Dye to perform PCR Quantitative real-time PCR - Bacterial DNA
Get tips on using Mastermix 16S Complete to perform PCR Quantitative real-time PCR - Bacterial DNA
Get tips on using jetPEI®-HUVEC to perform DNA transfection Mammalian cells - Primary cells HUVEC
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