siRNA / miRNA gene silencing Human Primary Endometrial Stromal Cells hsa-miR-542-3p

Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using KnockOut™ Serum Replacement - Multi-Species to perform Stem cell Differentiation media Human Limbal Epithelial cells

Get tips on using Anti-LC3 antibody produced in rabbit to perform Autophagy assay cell type - Human osteosarcoma cancer cells

Get tips on using PE Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - Human T-cells

Get tips on using FITC Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - Human T-cells

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using M-PER™ Mammalian Protein Extraction Reagent to perform Protein isolation Tissue - Human aortic endothelial cells

Get tips on using Quant-iT™ RiboGreen™ RNA Assay Kit to perform RNA quantification Fuorimetric - human trophoblast cells

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.