RNA sequencing Rat

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 PE-Cy™7 Rat Anti-Mouse TNF to perform Flow cytometry Anti-bodies Mouse - TNF-α

Get tips on using Alexa Fluor® 488 Rat Anti-Mouse CD146 to perform Flow cytometry Anti-bodies Mouse - CD146/MCAM

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - MIA PaCa-2

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - MDA-MB-231

Get tips on using PE-Cy™7 Rat anti-Mouse CD117 to perform Flow cytometry Anti-bodies Mouse - CD117/c-kit

Get tips on using PE-Cy™7 Rat Anti-Mouse CD31 to perform Flow cytometry Anti-bodies Mouse - CD31/Pecam-1