rna-isolation-purification-cells-primary-rat-cortical-neurons

Get tips on using Agencourt AMPure XP - PCR Purification to perform

Get tips on using Gibco™Neurobasal™ Medium to perform Stem cell culture media GBM patient-derived stem cells

Get tips on using SENSE mRNA-Seq Library Prep Kit V2 to perform RNA sequencing Rat - Hippocampal tissue

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells hsa-miR-542-3p Lipid

Get tips on using Gibco™Neurobasal™ Medium to perform Stem cell Differentiation media hiPSCs differentiation into Microglial-like cells

Get tips on using RNeasy 96 Kit (12) to perform mRNA / Ribonucleoprotein isolation / purification Ribonucleoprotein

Get tips on using Gibco™Neurobasal™-A Medium to perform Stem cell culture media GBM patient-derived stem cells

Get tips on using Subcellular Protein Fractionation Kit for Cultured Cells to perform Protein isolation Mammalian cells - Caco-2

Get tips on using Subcellular Protein Fractionation Kit for Cultured Cells to perform Protein isolation Mammalian cells - SH-SY5Y

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.