rna-isolation-purification-tissue-mouse-testes

Get tips on using EasySep™ Human Cord Blood CD34 Positive Selection Kit II to perform Cell Isolation CD34+ cells

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - Human eutopic endometrial stromal cells

Get tips on using EZ-10 Spin Column Plasmid DNA Miniprep Kit to perform Plasmid Isolation E. coli-S. cerevisiae transconjugate

Get tips on using EasySep™ Human B Cell Enrichment Kit II Without CD43 Depletion to perform Cell Isolation B cell

Get tips on using Lentivirus Rapid Quantitation Kit to perform RNA quantification Coloremetric

Get tips on using QuantiTect Virus Kit to perform RNA quantification qPCR

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.

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 RNeasy PowerClean Pro Cleanup Kit (50) to perform Removal of contamination in RNA Melanin contamination

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - H9