WebIf your vector has been cut with a single enzyme, you'll want to treat it with Calf Intestinal Alkaline Phosphatase (CIP) first. Set up the following reaction: * The amount of insert DNA used will depend on the relative size of the insert compared to the size of the vector. A molar ratio between insert and vector should be calculated. WebIt is essential if using crude extracts that protease inhibitors are included in the CIP buffer. 2. Spin down the samples. Re-suspend the pellets in the CIP buffer, 1 ug protein per 10 ul …
Alkaline Phosphatase Calf Intest (CIP), New England Biolabs
WebCIP is commonly used to remove 5’-phosphate groups from DNA, RNA and both ribo and deoxy-ribonucleoside triphosphates. Removal of 5’-phosphates is very useful in … WebCalf Intestinal Alkaline Phosphatase (CIAP) is a phosphomonoesterase that removes 3´ and 5´ phosphates from DNA and RNA. Concentration: 20 units/µL. Applications: Dephosphorylation of 5´-phosphorylated termini of … shypot plus s.r.o
Alkaline Phosphatase Calf Intest (CIP), New England …
WebMar 5, 2024 · Calf intestinal phosphatase (CIP) Catalyzes the removal of 5' phosphate groups from RNA, DNA and ribo- and deoxyribo- nucleoside triphosphates (e.g. ATP, rATP). CIP treated duplex DNA cannot self ligate. Hemi-phosphorylated duplexes will be ligated on one strand (the phosphorylated strand) and remain "nicked" on the other. WebDephosphorylation is a common step in traditional cloning workflows to ensure that the vector does not re-circularize during ligation. If a vector is linearized by a single restriction enzyme, or has been cut with two enzymes with compatible ends, use of a phosphatase, such as Quick CIP, to remove the 5´ phosphate reduces the occurrence of vector re … WebAug 3, 2024 · Alkaline Phosphatase Several types of alkaline phosphatases (or alkaline phosphomonoesterase) are commonly used in molecular cloning, including bacterial alkaline phosphatase (BAP) and calf intestinal alkaline phosphatase (CIP, CIAP, or CAP). Similar enzymes isolated from more esoteric cold-blooded organisms (e.g., SAP … shyplite international shipping