siRNA / miRNA gene silencing Rat Schwann cells

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.

Microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary rat aortic smooth muscle cells

Get tips on using Ad-Sal-shRNA to perform shRNA gene silencing Rat - H9c2 salusin-β

Get tips on using Ad-Sal-shRNA to perform shRNA gene silencing Rat - WKY Salusin-β

Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 GLUT1

Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 CD74

Get tips on using pSUPER.retro.neo+gfp vector- Syn G (exon 3) siRNA to perform shRNA gene silencing Mouse - RGC-5 Syn G (Exon 3)

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 ROCK2