Protein Expression Prokaryotic cells M. smegmatis

Get tips on using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814 to perform Immunohistochemistry Mouse - β-Catenin

Get tips on using CD279 (PD-1) Monoclonal Antibody (RMP1-30), FITC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD279/PD-1

Get tips on using CD273 (PD-L2) Antibody, anti-mouse, PerCP-Vio® 700 to perform Flow cytometry Anti-bodies Mouse - CD273/PD-L2

Get tips on using Pacific Blue™ anti-mouse Ly-6A/E (Sca-1) Antibody to perform Flow cytometry Anti-bodies Mouse - Ly-6A-E/Sca1

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

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

Get tips on using CD29 (Integrin beta 1) Monoclonal Antibody (eBioHMb1-1 (HMb1-1)), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD29/β1-Integrin

Get tips on using Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 568 conjugate to perform Flowcytometry Secondary Antibody - Goat Mouse Alexa Fluor 568

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. Multiplexing such a reaction amplifies the design challenges where one target requires 3 primers, which should be exclusively bound nowhere in the template DNA or to each other. Similarly, two targets require 6, three require 9, and so on. Each amplicon needs to be either a different size (for gels) or labeled with a different fluorescent tag that is spectrally distinct from the others in the reaction. Further complicating this, different targets in the reaction can compete with each other for resources and causes more challenges in the detection of amplicons. However, with proper primer designing, their validation, optimize quality and concentration of the enzyme and buffers certainly lead to a successful multiplex PCR reaction.

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?