CRISPR Mouse Deletion NIH 3T3

Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Mouse - Deletion 3T3-L1 SWELL1

Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Mouse - Deletion 3T3-L1 fmnl 2/3

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.

Get tips on using ON-TARGETplus Mouse Myb (17863) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - NIH-3T3 Myb

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.

Cell cycle can be challenging due to difference introduced by sample handling, timing, and difference within the sample. Downstream instriuments to analyse cell cycle (Multicolor flow cytometry and multicolor imaging) can answer these challenges. Relevant markers can be combined with cell phenotyping markers to look at events within subpopulations of cells.

Get tips on using Purified Mouse Anti-Beclin Clone 20/Beclin (RUO) to perform Autophagy assay cell type - NIH-3T3

Get tips on using QuantiFluor® dsDNA System to perform DNA quantification Mouse - NIH 3T3

Get tips on using SMS2 CRISPR/Cas9 KO Plasmid (m) to perform CRISPR Mouse - Deletion C2C12 Sgms2

Get tips on using SMS1 CRISPR/Cas9 KO Plasmid (m) to perform CRISPR Mouse - Deletion C2C12 Sgms1