rna-isolation-purification-cells-immortalized-ags

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Trichloroacetic acid to perform Protein isolation Mammalian cells - Human gingival epithelial cells

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - Human CD14+ cells

Get tips on using Agilent RNA 6000 Pico Kit to perform RNA quantification Fuorimetric

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Coloremetric

Get tips on using Qubit RNA BR Assay Kit to perform RNA quantification Coloremetric

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - HEK293

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - HeLa

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Bacteria - Gram positive piezophilic bacteria [AT7 and AT12 Strains]