Get tips on using Ad-Sal-shRNA to perform shRNA gene silencing Rat - H9c2 salusin-β
Get tips on using Ad-Sal-shRNA to perform shRNA gene silencing Rat - WKY Salusin-β
Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - HEK293 human embryonic kidney cells
Get tips on using pRNAT-H1.1/Neo to perform shRNA gene silencing Mouse - 4T1 Integrin α6
Get tips on using DMEM/F12 - Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 to perform Stem cell culture media Ovarian cancer stem cells (Caov3, 3AO, SKOV3)
Get tips on using GeneSilencer® Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - C6 Cationic lipid based
Get tips on using Individual: TRC Mouse Cdh1 shRNA to perform shRNA gene silencing Mouse - 4T1 Cdh1
Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.
Reporter gene assays are designed to test the regulation of the expression of a gene of interest. This is usually done by linking the promoter of the gene of interest with a gene such as a firefly luciferase, which can be easily detected by addition of luciferin that leads to an enzymatic reaction to produce luminescence. The enzymatic reaction can be correlated to the expression of the gene of interest. Another luciferase gene that can be used is Renilla luciferase. For an appropriate luciferase assay: 1. the reporter should express uniformly in all cells, 2. specifically respond to effectors that the assay intends to monitor, 3. have low intrinsic stability to quickly reflect transcriptional dynamics. It is important to have an equal number of cells plated in each testing condition to avoid any incorrect readouts. Reporter assays could be single or dual reporter assays. The reporter could be both luciferases. Most dual-luciferase assays involve adding two reagents to each sample and measuring luminescence following each addition. Adding the first reagent activates the first luciferase reporter reaction; adding the second reagent extinguishes first luciferase reporter activity and initiates the second luciferase reaction. Dual-luciferase assays have some advantages, including 1. reduces variability, 2. reduces background, 3. normalizes differences in transfection efficiencies between samples.
Get tips on using EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation Kit to perform ChIP Human - Fibroblast cell lines
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