siRNA / miRNA gene silencing Human HNSCC cell line

Get tips on using AllPrep DNA/RNA/miRNA Universal Kit to perform RNA isolation / purification Tissue - Mouse Adipose

Get tips on using AllPrep DNA/RNA/miRNA Universal Kit to perform RNA isolation / purification Tissue - mouse adipose tissue

Get tips on using Type-it Fast SNP Probe PCR Kit (4000) to perform Cell line authentication Peripheral blood lymphocytes

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 pRNAT-H1.1/Neo to perform shRNA gene silencing Mouse - 4T1 Integrin α6

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

Get tips on using Galacto-Star™ β-Galactosidase Reporter Gene Assay System for Mammalian Cells to perform Reporter gene assay β-galactosidase substrates - MCF-7 human breast cancer

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.