siRNA / miRNA gene silencing Human HeLa

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 Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram negative Helicobacter pylori

Get tips on using pLX-sgRNA to perform CRISPR Human - Deletion CIITA

Get tips on using MethylEasy™ Xceed (ME002) to perform DNA methylation profiling Gene specific profiling - Rat whole pituitary glands PROP1

Get tips on using EZ DNA Methylation-Direct Kit to perform DNA methylation profiling Gene specific profiling - A2780 miR-30a-5p

Get tips on using EZ DNA Methylation-Direct Kit to perform DNA methylation profiling Gene specific profiling - A2780 miR-30c-5p

Get tips on using CpGenome Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - Ca Ski HPV-16

Get tips on using Imprint® Methylated DNA Quantification Kit to perform DNA methylation profiling Gene specific profiling - MRC-5 LSH

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - rat nucleus pulposus

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - adipose stem cells