Site Directed Mutagenesis (SDM) Human Point mutation THP-1

Get tips on using Tlr4 Mouse siRNA Oligo Duplex to perform siRNA / miRNA gene silencing Mouse - BV2 TLR4

Get tips on using ON-TARGETplus Mouse Lgals3 siRNA to perform siRNA / miRNA gene silencing Mouse - BV2 LGAL3S3

Get tips on using Cox-2 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - B16-F10 COX-2

Get tips on using ON-TARGETplus Mouse Stat3 siRNA to perform siRNA / miRNA gene silencing Mouse - RAW264.7 STAT3

Get tips on using ON-TARGETplus Mouse Prkaa1 siRNA to perform siRNA / miRNA gene silencing Mouse - RAW264.7 Prkaa1

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 AM5770: pSilencer™ 3.1-H1 neo to perform shRNA gene silencing Mouse - CT26 OPN

Get tips on using IL-4Rα shRNA (m) Lentiviral Particles to perform shRNA gene silencing Mouse - R221a IL4Rα

Get tips on using IGF-IRα/β siRNA (r) to perform siRNA / miRNA gene silencing Rat - RGC-5 IGF1R

Get tips on using ON-TARGETplus Rat Fyn siRNA to perform siRNA / miRNA gene silencing Rat - Schwann cells Fyn