Site Directed Mutagenesis (SDM) Human Point mutation

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 ON-TARGETplus Human BSG / emmprin (682) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - BCP-1 Emmprin / BSG

Get tips on using Human PLZF Antibody to perform Immunohistochemistry Mouse - PLZF

Get tips on using Human SOX9 Antibody to perform Immunohistochemistry Mouse - SOX9

Get tips on using Human TAK1 Antibody to perform Western blotting Tak1

Get tips on using Human VEGF ELISA Kit (ab100662) to perform ELISA Human - VEGF

Get tips on using Human VEGF 165b DuoSet ELISA to perform ELISA Human - VEGF

Get tips on using Human RBP4 Quantikine ELISA Kit to perform ELISA Human - RBP4

Get tips on using Human Prolactin/PRL ELISA Kit to perform ELISA Human - PRL

Get tips on using Human Osteopontin ELISA Kit (ab100618) to perform ELISA Human - OPN