Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines OVCAR-3
Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines SH-SY5Y
Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer
Get tips on using High Pure PCR Product Purification Kit to perform DNA gel extraction / PCR product purification Product size > 15Kb
Get tips on using Cell Proliferation Reagent WST-1 to perform Cell cytotoxicity / Proliferation assay cell type - SMMC-7721, Huh7, Hep3B, 293T
Get tips on using ApoAlert™ DNA Fragmentation Assay Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer
Get tips on using mirVana® miRNA mimic to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells hsa-miR-542-3p
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.
Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,
Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,
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