RNA sequencing

Get tips on using Endothelin 1 ELISA Kit (ab133030) to perform ELISA Rat - Endothelin 1

Get tips on using BMP-2 Quantikine ELISA Kit to perform ELISA Rat - BMP-2

Get tips on using MicroRNA isolation kit to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR

Get tips on using CD38 CRISPR Activation Plasmid (r) to perform CRISPR Rat - Activation CD38

Get tips on using ATG7 antibody [N3C2], Internal to perform Autophagy assay cell type - RAW 264.7

Get tips on using LAMP-1 Polyclonal Antibody to perform Autophagy assay cell type - RAW 264.7

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Primary cells Rat astrocytes

Get tips on using FITC BrdU Flow Kit to perform Cell cycle assay mouse - RAW 264.7

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 Recombinant Anti-SOX9 antibody [EPR14335] (ab185230) to perform Immunohistochemistry Rat - Sox9