shRNA gene silencing Mouse Prostate cancer cell lines (DU145 and PC3) CD24

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using APC anti-mouse CD140a Antibody to perform Flow cytometry Anti-bodies Mouse - CD140/PDGFR-α

Get tips on using PE Rat Anti-Mouse CD140A to perform Flow cytometry Anti-bodies Mouse - CD140/PDGFR-α

Get tips on using - to perform siRNA / miRNA gene silencing Rat - IEC-6

Get tips on using Fenozol to perform siRNA / miRNA gene silencing Human - BOSC23

Get tips on using ON-TARGETplus Human PCSK7 (9159) siRNA - Individual to perform siRNA / miRNA gene silencing Human - A253 IGFBP-7

Get tips on using PE Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - T-cells Mouse (CD4+ and CD8+)

Get tips on using FuGENE® HD Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines PC12

Get tips on using p53 siRNA to perform siRNA / miRNA gene silencing Human - COV-434 p53

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.