RNA isolation / purification Plants

Get tips on using RNAprotect Bacteria Reagent to perform RNA stabilization Microbial

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 Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 HER2 leader peptide

Get tips on using NucleoSpin® Gel and PCR Clean-up to perform DNA gel extraction / PCR product purification Product size > 15Kb

Get tips on using Ni-NTA Magnetic Agarose Beads (6 x 1 ml) to perform Protein tag Purification of His-tagged proteins

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 BioMag Goat Anti-Rat IgG (500 ml) to perform Cell Isolation Mouse T cells

Get tips on using Trichloroacetic acid to perform Protein isolation Mammalian cells - Human gingival epithelial cells

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - THP-1

Get tips on using RIPA Lysis Buffer (ProteinSimple) to perform Protein isolation Tissue - Mouse cardiac tissue