dna-quantification-human-hela

Get tips on using Hot StarTaq Master Mix Kit (2500 U) to perform PCR Hot start PCR - Mammalian DNA

Get tips on using QuantiTect SYBR Green RT-PCR Kit (1000) to perform PCR Conventional / Qualitative PCR - mammalian DNA

Get tips on using EpiTect MethyLight PCR +ROX Vial Kit (200) to perform PCR Methylation specific PCR - Mammalian DNA

Get tips on using illustra tissue and cells genomicPrep Mini Spin Kit to perform DNA isolation / purification Tissue - kidney

Get tips on using GenLadder 50bp (ready-to-use) with dye Orange G to perform DNA Ladder 50 bp

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.

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 Silencer® Select GLO-1 siRNA to perform siRNA / miRNA gene silencing Human - Primary Human Aortic Endothelial Cells GLO-1 Lipid

Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - HNSCC Detroit 562 human head and neck tumor cells

Get tips on using AmpFLSTR™ Identifiler™ Direct PCR Amplification Kit to perform Cell line authentication Human iPSC cells derived from human dermal fibroblasts