siRNA / miRNA gene silencing Mouse NIH-3T3

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.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using RORC siRNA-NM_011281 to perform siRNA / miRNA gene silencing Mouse - FL83B Rorc

Get tips on using Mm_Pttg1ip_7 FlexiTube siRNA to perform siRNA / miRNA gene silencing Mouse - AtT20 PTTG1

Get tips on using Mm_Gadd45b_4 FlexiTube siRNA to perform siRNA / miRNA gene silencing Mouse - AtT20 GADD45β

Get tips on using Cdx2 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - mESC Cdx2

Get tips on using BECN1 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - BV2 BECN1

Get tips on using CD98 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - Colon26 CD98

Get tips on using CD98 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - RAW264.7 CD98

Get tips on using Mm_Hdac5_2 FlexiTube siRNA to perform siRNA / miRNA gene silencing Mouse - RAW264.7 HDAC5