siRNA / miRNA gene silencing Mouse B16 Melanoma 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.

Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform Microarray Gene expression arrays - Human endometrial stromal cells Biotin

Get tips on using Cellular Senescence Staining Kit to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Get tips on using psiCHECK-2vector to perform Reporter gene assay luciferase - HEK 293 human embryonic kidney cells

Get tips on using ElectraSense 4x2K/12K to perform Microarray Gene expression arrays - Human whole blood cells Biotin

Get tips on using pGL3-Basic Vector to perform Reporter gene assay luciferase - vascular smooth muscle cells (VSMC)

Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA methylation profiling Whole genome profiling - C2C12 mouse myoblast cells

Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. 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 the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - HT22 mouse hippocampal cells