Get tips on using DMEM/F-12 to perform 3D Cell Culture Media Mouse primary breast cancer ephitelial cells-Mammospheres
Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized SK-BR-3
Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized MDA-MB-231
Get tips on using miRcute miRNA Isolation Kit to perform RNA isolation / purification Cells - immortalized MDA-MB-157
Get tips on using HiPurA™ Total RNA Miniprep Purification Kit to perform RNA isolation / purification Cells - immortalized HaCaT
Get tips on using Maxwell® 16 LEV simplyRNA Purification Kit to perform RNA isolation / purification Cells - immortalized A549
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 PicoPure™ RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized U-251
Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized Saos-2
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