RNA sequencing Human

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 PE Mouse Anti-Human CD140a to perform Flow cytometry Anti-bodies Human - CD140/PDFGR2

Get tips on using PE Mouse Anti-Human CD184 to perform Flow cytometry Anti-bodies Human - CD184/CXCR4

Get tips on using APC Mouse Anti-Human CD184 to perform Flow cytometry Anti-bodies Human - CD184/CXCR4

Get tips on using FITC Mouse Anti-Human CD36 to perform Flow cytometry Anti-bodies Human - CD36/CB38

Get tips on using Purified Mouse Anti-Human CD36 to perform Flow cytometry Anti-bodies Human - CD36/CB38

Get tips on using ON-TARGETplus human ATG16L1 siRNA to perform siRNA / miRNA gene silencing Human - SHSY5Y ATG16L1

Get tips on using MISSION® esiRNA_ human CCL2 to perform siRNA / miRNA gene silencing Human - U251 CCL2

Get tips on using ON-TARGETplus Human TET1 siRNA to perform siRNA / miRNA gene silencing Human - A172 TET1

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.