DNA gel extraction / PCR product purification

Get tips on using Qproteome Bacterial Protein Prep Kit to perform Protein isolation Bacteria - Salmonella paratyphi A

Get tips on using pFRT to perform Protein Expression Eukaryotic cells - HeLa R19 cMBL

Get tips on using pGreen-TRAIL to perform Protein Expression Eukaryotic cells - N. benthamiana TRAIL

Get tips on using pVRB2B3 to perform Protein Expression Eukaryotic cells - HEK293 hβ-defensin 2/3

Get tips on using pHR-CMV-TetO2-CD45 to perform Protein Expression Eukaryotic cells - HEK293 CD45

Get tips on using pHR-CMV-TetO2-VSV-G to perform Protein Expression Eukaryotic cells - HEK293 VSV-G

Get tips on using pSpCas9(BB)-2A-Puro (PX459) to perform CRISPR Mouse - Deletion ES (embryonic stem) cells Etv2 promoter

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 pTY- α-amylase to perform Protein Expression Prokaryotic cells - E. coli Pyrococcus woesei Hyperthermophile α-Amylase

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.