DNA gel extraction / PCR product purification

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat pulmonary artery smooth muscle cell (pPASMC)

Get tips on using FuGENE® HD Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human pulmonary artery smooth muscle cells (HPASMC)

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human pulmonary artery smooth muscle cells (HPASMC)

Get tips on using APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU to perform DNA Damage Assay Human Skin Fibroblast Cell (FSK)

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - Cancer cell lines Liver cancer cell lines Hepato cellular carcenoma (SMMC-7721, Huh7 & HepG2))

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - TCP-1, BCPAP & nthy-ori 3-1 (thyroid tumor cells) METTL7A

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - TCP-1, BCPAP & nthy-ori 3-1 (thyroid tumor cells) BCPAP

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.

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.