Get tips on using Pierce™ Cell Surface Protein Isolation Kit to perform Protein isolation Mammalian cells - Human aortic endothelial cells
Get tips on using Silencer® FANCD2 siRNA (human) to perform siRNA / miRNA gene silencing Human - 501 Mel and SK Mel 28 FANCD2
Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb to perform Cell cytotoxicity / Proliferation assay cell type - K562
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.
Get tips on using Dansylcadaverine to perform Autophagy assay cell type - MG-63
Get tips on using LY294002 to perform Autophagy assay cell type - SH-SY5Y
Get tips on using Diogenes to perform ROS assay cell type - CHO-K1
Get tips on using H4A3 to perform Autophagy assay cell type - SH-SY5Y
Get tips on using p62 (human) polyclonal antibody to perform Immunohistochemistry Mouse - p62
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