Acid phosphatase detection heavily relies on determining the concentration of tartrate-resistant acid phosphatase (TRAP) in the sample. Hence, sample preparation is very crucial and it should be done strictly as per kit manufacturer instructions to avoid any inconsistency and poor sensitivity.
Get tips on using PAXgene Tissue miRNA Kit to perform RNA isolation / purification Tissue - rat brain tissue
Get tips on using PAXgene Tissue miRNA Kit to perform RNA isolation / purification Tissue - rat spleen tissue
Get tips on using PAXgene Tissue miRNA Kit to perform RNA isolation / purification Tissue - rat heart muscle tissue
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 TRIzol Reagent to perform RNA isolation / purification Cells - primary human mononuclear cells
Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human endothelial cells
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