PCR

Get tips on using pET30a-β4 to perform Protein Expression Prokaryotic cells - E. coli E. granulosus β4 tubulin

Get tips on using pET30a-α9 to perform Protein Expression Prokaryotic cells - E. coli E. granulosus α9 tubulin

Get tips on using Beclin-1 Antibody, ProSci to perform Autophagy assay cell type - IEC-6

Get tips on using EpiTect MSP Kit to perform DNA methylation profiling Gene specific profiling - HeLa MEG3

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - HepG2 Fibrinogen

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Gene specific profiling - SKOV3 AGR2

Get tips on using EZ4U - Cell Proliferation Assay to perform Live / Dead assay mammalian cells - GH3

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Whole genome profiling - mouse iPSCs

Get tips on using CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) to perform Cell cytotoxicity / Proliferation assay cell type - LTEP-a-2 lung adenocarcenoma

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.