rna-isolation-purification-tissue-mouse-spleen

Get tips on using Fenozol to perform RNA isolation / purification Tissue - human aorta tissue

Get tips on using RNeasy Lipid Tissue Mini Kit to perform RNA isolation / purification Cells - primary human brain microvascular endothelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - rat brain tissue

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - rat heart tissue

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - rat lung tissue

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - rat liver tissue

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Tissue - rat pancreas tissue

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Rat Pineal gland

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.

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.