siRNA / miRNA gene silencing Human Min-6

Get tips on using TruSeq ChIP Library Preparation Kit to perform ChIP Human - PBMC

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Human - LCL

Get tips on using TruSeq ChIP Library Preparation Kit to perform ChIP Human - HUVEC

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Human - HepG2

Get tips on using Pierce™ Agarose ChIP Kit to perform ChIP Human - HeLa

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Human - T47D

Get tips on using HSP70 High Sensitivity ELISA kit to perform ELISA Human - HSP70

Get tips on using Mouse GDNF ELISA Kit (ab171178) to perform ELISA Human - GDNF

Get tips on using TruSeq RNA Library Prep Kit v2 to perform RNA sequencing Human - MIA PaCa-2

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.