Site Directed Mutagenesis (SDM) Human Insertion IMR-32

Get tips on using REG1 beta Polyclonal Antibody to perform Immunohistochemistry Human - REG1

Get tips on using Anti-Hes1 antibody (ab49170) to perform Immunohistochemistry Human - Hes1

Get tips on using Anti-Notch1 antibody (ab27526) to perform Immunohistochemistry Human - Notch1

Get tips on using MLH1 antibody [G168-15] to perform Immunohistochemistry Human - MLH1

Get tips on using Anti-CRISP3 antibody (ab105951) to perform Immunohistochemistry Human - CRISP3

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Human - RCC

Get tips on using MAGnify™ Chromatin Immunoprecipitation System to perform ChIP Human - OSCC

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

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.

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.