DNA methylation profiling Gene specific profiling Human ovarian tissue

Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA quantification Human - WI-38

Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA quantification Human - HEK 293

Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA quantification Human - THP 1

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - human breast tissue

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - human lung tissue

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - human placental tissue

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - human brain tissue

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - human aorta tissue

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.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.