rna-isolation-purification-cells-immortalized-em-2

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using MAP LC3β Antibody (G-2) to perform Autophagy assay cell type - AR42J

Get tips on using ErbB2 (HER-2) Antibody Cocktail to perform Western blotting HER2

Get tips on using BLOOD AGAR BASE NO.2 to perform Bacterial cell culture media Helicobacter pylori

Get tips on using EpiTect 96 Bisulfite Kit (2) to perform Bisulfite DNA Modification Fluids

Get tips on using FaqI (BsmFI) (2 U/µL) to perform Restriction Enzymes BsmFI / FaqI

Get tips on using BspPI (AlwI) (2 U/µL) to perform Restriction Enzymes AlwI / BspPI

Get tips on using AllPrep Bacterial DNA/RNA/Protein Kit (50) to perform DNA isolation / purification Bacteria - Gram negative Massilia sp

Get tips on using TIMP-2 Antibody (B-12): sc-365671 to perform Western blotting TIMP-2