Protein Expression Prokaryotic cells S. lividans

Get tips on using Nuclear Extract Kit to perform Protein isolation Mammalian cells - HOG

Get tips on using RIPA Buffer (10X) to perform Protein isolation Mammalian cells - Huh7

Get tips on using Mammalian Cell Lysis kit to perform Protein isolation Mammalian cells - STTG1

Get tips on using RIPA Lysis Buffer System to perform Protein isolation Mammalian cells - HOG

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - Caco-2

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - BHK-21

Get tips on using RIPA Lysis Buffer, 10X to perform Protein isolation Mammalian cells - STTG1

Get tips on using 2-D Quant Kit to perform Protein quantification Mammalian cells - SiHa

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.