Site Directed Mutagenesis (SDM) Human Point mutation THP-1

Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)

Get tips on using Gibco™Ham's F-12K (Kaighn's) Medium to perform Stem cell culture media Human myogenic progenitor cells

Get tips on using SMARTpool: ON-TARGETplus TP63 siRNA to perform siRNA / miRNA gene silencing Human - A253 P36

Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform siRNA / miRNA gene silencing Human - HT-1376 GLUT1

Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform siRNA / miRNA gene silencing Human - HT-1376 CD74

Get tips on using Hs_TET3_2 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - HCT-116 TET3(TET methylcytosine dioxygenase 3)

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - A549 human adenocarcinomic human alveolar basal epithelial cells

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform siRNA / miRNA gene silencing Human - HT-1376 ROCK2

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.