Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase
Get tips on using siGENOME Human MINK1 siRNA to perform siRNA / miRNA gene silencing Human - RMS MINK
Get tips on using siGENOME Human MAP4K2 siRNA to perform siRNA / miRNA gene silencing Human - RMS MAP4K2
Get tips on using esiRNA human PTPN3 (esiRNA1) to perform siRNA / miRNA gene silencing Human - A2780 PTPN3
Get tips on using siGENOME Human PAK1 siRNA to perform siRNA / miRNA gene silencing Human - HeLa PAK1
Get tips on using ON-TARGETplus SMARTpool - Human to perform siRNA / miRNA gene silencing Human - U2OS DKC1
Get tips on using Anti-trimethyl-Histone H3 (Lys9) Antibody to perform ChIP H3K9me3 - Sheep Rat -NA-
Get tips on using Anti-trimethyl-Histone H3 (Lys4) Antibody to perform ChIP H3K4Me3 - Sheep Rat -NA-
Get tips on using truChIP Ultra-Low Chromatin Shearing Kit with Formaldehyde to perform ChIP Mouse - Osteoblasts
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.
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