dna-quantification-human-pc-3

Get tips on using Ion Total RNA-Seq Kit v2 to perform RNA sequencing Rat - PC12

Get tips on using TruSeq RNA Library Prep Kit v2 to perform RNA sequencing Rat - PC12

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Mouse - Deletion B16-F1 PC7

Get tips on using CM-H2DCFDA (General Oxidative Stress Indicator) to perform ROS assay cell type - PC12

Get tips on using MYH9 CRISPR/Cas9 KO Plasmid (h) to perform CRISPR Rat - Deletion PC12 myosin IIA (Myh9)

Get tips on using ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - PC12 Atf4

Get tips on using OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - PC12

Get tips on using Monoclonal Anti-Proliferating Cell Nuclear Antigen antibody produced in mouse to perform Western blotting PCNA

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.

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.