crispr-human-repression-akt2

Get tips on using ApopTag® Peroxidase In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - HNSCC Detroit 562 human head and neck tumor cells

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Human - Precision cut lung slices Target preparation kit (RNA Amplification + Hybridization + control)

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using GenomONE™-Neo HVJ-E Membrane Fusion Transfection Kit to perform siRNA / miRNA gene silencing Human - U937 MK2 (MAPK Kinase 2) Viral vectors

Get tips on using GenomONE™-Neo HVJ-E Membrane Fusion Transfection Kit to perform siRNA / miRNA gene silencing Human - Jurkat MK2 (MAPK Kinase 2) Viral vectors

Get tips on using Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay to perform TUNEL assay cell type - A549, NCI-H460, H1299 human alveolar carcinoma

Get tips on using Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using Agilent DNA 1000 Kit Bioanalyzer DNA Analysis Part Number:5067-1504 to perform Cell line authentication Human lung carcinoma cell line NCI-H1299

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)

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. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.