shRNA gene silencing Human Islets of langerhans

Get tips on using ON-TARGETplus Mouse Myb (17863) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - NIH-3T3 Myb

Get tips on using ON-TARGETplus Mouse Ddit4 (74747) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RGC-5 Ddit4

Get tips on using ON-TARGETplus Mouse Eif2ak3 (13666) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 Perk/Eif2ak3

Get tips on using ON-TARGETplus Mouse Casp8 (12370) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 caspase-8

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.

Get tips on using Flot1 Rat siRNA Oligo Duplex (Locus ID 64665) to perform siRNA / miRNA gene silencing Rat - NRCM Flot1

Get tips on using Flot2 Rat siRNA Oligo Duplex (Locus ID 83764) to perform siRNA / miRNA gene silencing Rat - NRCM Flot2

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.