Incubate 60 minutes at … Add 0.5 - 1.0 unit/ug protein. Protocol for Dephosphorylation of 5'-ends of DNA using CIP (NEB #M0290) Usage Guidelines. Quick CIP is also active in 1X NEBuffers 1.1, 2.1, 3.1 as well as NEBuffers 1, 2, 3 and 4 and NEBuffer for EcoRI. 3. 240 County Road Ipswich, MA 01938-2723 978-927-5054 (Toll Free) 1-800-632-5227 Fax: 978-921-1350 Info@neb.com Protocol for Dephosphorylation of 5´-ends of DNA using CIP (NEB #M0290) Protocols.io also provides an interactive version of this protocol where you can discover and share optimizations with the research community.. Protocol. Add protien/lysate to 1x NEB buffer (0.5-1.0 ug/10ul) 2.
Quick CIP is a heat-labile version of calf intestinal alkaline phosphatase (CIP) purified from a recombinant source. Chromatin immunoprecipitation (ChIP) reveals the dynamics of histone modifications and transcription co-factors, bound to specific genes, or their binding in genome-wide profiles using microarray or sequencing (1). Exo-CIP Rapid PCR Cleanup Kit enables consistent read lengths for amplicons of varying length PCR products of varying lengths were treated with Exo-CIP Rapid PCR Cleanup Kit or ExoSAP-IT PCR Cleanup Reagent following manufacturer-recommended protocols (5 min vs. 30 min). Some protocols also recommend the addition of more CIP for these difficult ends. Protocol for Dephosphorylation of 5´-ends of DNA using CIP in Restriction Enzyme Reaction. Prepare a 20 μl reaction as follows: EDTA), inorganic phosphate and phosphate analogs. ChIP data always represents a population of nucleosomes, from cultured cells or tissues.
Quick CIP is inhibited by metal chelators (e.g. We have found that the difference in efficiency is about 20-25%. Quick CIP activity is enhanced in the presence of monovalent salts. Dephosphorylating Proteins with CIP: CIP can be used to release phosphate groups from phosphorylated tyrosine, serine and threonine residues in proteins, using the following protocol: 1. ... Why do some protocols recommend using CIP at 50°C and some recommend 37°C?
CIP works slightly better on 3´ extensions and blunt-ends at 50°C. Uses the Calf Intestinal Alkaline Phosphatase (CIP - M0290) What phosphate groups are removed by rSAP, Quick CIP, or Antarctic Phosphatase (AnP)?