siRNA / RNAi /miRNA transfection Rat A-10

Get tips on using Anti-Collagen, Type VII to perform Immunohistochemistry Collagen Type VII - Rabbit Human -NA-

Get tips on using 53BP1 Antibody (H-300) to perform TissueFAxs 53BP1 [H-300] - Rabbit Human -NA-

Get tips on using 53BP1 Antibody (H-300) to perform Immunofluorscence  53BP1 [H-300] - Rabbit Human -NA-

Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Mammalian cells - RAW264.7

Get tips on using Pierce™ BCA Protein Assay Kit to perform Protein quantification Mammalian cells - RAW264.7

Get tips on using Ki-67 Antigen, Clone MIB-1 to perform Immunohistochemistry Ki67 - Rabbit Mouse / Human -NA-

Get tips on using Pierce™ Coomassie (Bradford) Protein Assay Kit to perform Protein quantification Mammalian cells - RAW264.7

Get tips on using Cultrex® BME Cell Invasion Assay to perform Cell migration / Invasion cell type - RAW 264.7

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.