Site Directed Mutagenesis (SDM) Rat Point mutation H9C2

Get tips on using Stealth siRNA(h)_GATA1 to perform siRNA / miRNA gene silencing Human - LAD2 GATA1

Get tips on using Stealth siRNA(m)_Atg5 to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Atg5

Get tips on using Stealth siRNA™ NOTCH2 to perform siRNA / miRNA gene silencing Human - THP-1 NOTCH2

Get tips on using Stealth siRNA™ NOTCH1 to perform siRNA / miRNA gene silencing Human - THP-1 NOTCH1

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

Get tips on using Gibco™ StemPro™ hESC SFM to perform Stem cell Differentiation media hiPSCs or hESCs differentiation to Embryoid body (EB)

Get tips on using Stealth siRNA(m) Atg16l2 to perform siRNA / miRNA gene silencing Mouse - Pancreatic Acinar cells Atg16l2

Get tips on using stealth siRNA GIRK1/KCNJ3 to perform siRNA / miRNA gene silencing Human - MDA-MB-453 GIRK1/KCNJ3

Get tips on using stealth siRNA GIRK1/KCNJ3 to perform siRNA / miRNA gene silencing Human - MDA-MB-231 GIRK1/KCNJ3

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.