DNA methylation profiling Gene specific profiling TCP-1, BCPAP

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

Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10

Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram negative Enterobacteriaceae

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram positive Mycobacterium tuberculosis

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram positive Lactobacillus amylovorus

Get tips on using T4 DNA Ligase (5 U/µL) to perform DNA ligation

Get tips on using PowerSoil® DNA isolation to perform AAA for reviews

Get tips on using AllPrep DNA/RNA Mini Kit to perform RNA isolation / purification Cells - immortalized Hepa-1c1c7

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.