rna-isolation-purification-cells-immortalized-bxpc-3

Get tips on using Attractene Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines OVCAR-3

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines BXPC3

Get tips on using DNA Isolation Kit for Cells and Tissues to perform DNA isolation / purification Cells - Primary cells Human primary keratinocytes

Get tips on using Total Exosome RNA & Protein Isolation Kit to perform Protein isolation Mammalian cells - HeLa

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.

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines SKOV-3

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.