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

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes

Get tips on using IGEPAL® CA-630 to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Cells - primary human osteoblasts

Get tips on using NanoOrange™ Protein Quantitation Kit to perform Protein quantification Mammalian cells - Human podocytes

Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - human trophoblast cells

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Human - Endometrial stromal cells Target preparation kit (Amplification + Hybridization + control)

Get tips on using EpiQuik Dnmt3A Assay Kit to perform DNA methylation profiling Whole genome profiling - human peripheral blood mononuclear 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.

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.

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.