3D Cell Culture Media

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 BrdU Cell Proliferation Assay to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma

Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma

Get tips on using Cell Proliferation ELISA, BrdU to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma

Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - LTEP-a-2 lung adenocarcenoma

Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - SMMC-7721, Huh7, Hep3B, 293T

Get tips on using Cell DNA Isolation Kit to perform DNA isolation / purification Cells - Immortalized cell lines C2C12

Get tips on using Cell Death Detection ELISA to perform Apoptosis assay cell type - Human endometrial stromal cells

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Gentra Puregene Cell Kit to perform DNA isolation / purification Cells - Immortalized cell lines SH-SY5Y