shRNA gene silencing Rat H9c2

Get tips on using Color-coded Prestained Protein Marker, High Range (43-315 kDa) #12949 to perform Protein Ladder Prestained

Get tips on using Color-coded Prestained Protein Marker, Broad Range (10-250 kDa) #74124 to perform Protein Ladder Prestained

Get tips on using Prestained Protein Ladder – Mid-range molecular weight (10 - 180 kDa) (ab116027) to perform Protein Ladder Prestained

Get tips on using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb #9728 to perform ChIP Anti-bodies H3K27me2

Get tips on using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb #5326 to perform ChIP Anti-bodies H3K4me1

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - BxPc-3 human primary pancreatic adenocarcinoma

Get tips on using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814 to perform Immunohistochemistry Mouse - β-Catenin

Get tips on using CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) to perform Cell cytotoxicity / Proliferation assay cell type - MG-63

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.