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 EasySep™ Human B Cell Enrichment Kit to perform Cell Isolation B cell
Get tips on using RosetteSep™ Human B Cell Enrichment Cocktail to perform Cell Isolation B cell
Get tips on using EasySep™ Human B Cell Isolation Kit to perform Cell Isolation B cell
Get tips on using StemSep™ Human CD34 Positive Selection Cocktail to perform Cell Isolation CD34+ cells
The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.
Get tips on using Brilliant Violet 510™ anti-human HLA-DR Antibody to perform Flow cytometry Anti-bodies Human - HLA-DR
Get tips on using PerCP-Cy™5.5 Mouse Anti-Human HLA-DR to perform Flow cytometry Anti-bodies Human - HLA-DR
Get tips on using PE/Dazzle™ 594 anti-human CD184 (CXCR4) Antibody to perform Flow cytometry Anti-bodies Human - CD184/CXCR4
A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.
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