DNA methylation profiling Gene specific profiling TCP-1, BCPAP

Get tips on using GeneJuice® Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines 3T3-L1

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid-mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time-consuming, but provide a more permanent expression of RNAi in the cells and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines.

Get tips on using DNeasy PowerBiofilm Kit (50) to perform DNA isolation / purification Biofilm samples

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - adipose

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - placenta

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - eye

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - lung

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - brain

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - THP-1

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.