rna-isolation-purification-tissue-rat-sublingual-glands

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples 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 MicroRNA isolation kit to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR

Get tips on using Biotin Rat Anti-Mouse OX40 Ligand (CD252) to perform Flow cytometry Anti-bodies Mouse - CD252/OX40L

Get tips on using PE-CF594 Rat Anti-Mouse Siglec-F to perform Flow cytometry Anti-bodies Mouse - Siglec F

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 ISOGEN to perform RNA isolation / purification Cells - primary human epidermal melanocytes

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Streptococcus pyogenes

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Staphylococcus epidermidis

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Listeria monocytogens

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum