shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y) Connexin 43

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

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 Monoclonal Mouse Anti-Human Ki-67 Antigen (Dako Omnis) Clone MIB-1 to perform Immunohistochemistry Human - Ki-67

Get tips on using Beta-Glo® Assay System to perform Reporter gene assay β-galactosidase substrates - rat mesenchymal stem cells (MSCs)

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - mouse pancreatic stellate cells

Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

Get tips on using ONE-Glo™ Luciferase Assay System to perform Reporter gene assay luciferase - BHK-21 baby hamster kidney cells

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.