Life science protocols and methods » Cloning and subcloning » DNA dephosphorylation
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| DNA dephosphorylation - http://dna.dephosphorylation Vector dephosphorylation step Dephosphorylation step is necessary to prevent self-ligation of cloning vectors in blunt-end and single restriction enzyme cloning. Calf intestinal alkaline phosphatase (CIAP) is most frequently used to remove 5' phosphate groups from cloning vector. To calculate the amount pmol of ends for linear dsDNA, use equation bellow. (ug DNA/(kb size of DNA))*3.04 |
| DNA Dephosphorylation protocols - MyBio - http://mybio.wikia.com/wiki/DNA_Dephosphorylation_protocols Apr 3, 2008 ... Includes a protocol for ligation. Dephosphorylation of DNA ends Dephosphorylation of DNA ends using Shrimp Alkaline Phosphatase (also known ... |
| Dephosphorylation of linearised plasmid DNA for ligation - http://users.path.ox.ac.uk/~ciu/Methods/DNA/dephosphorylation.html Dephosphorylation of linearised plasmids is a method employed to prevent the ... Extract the DNA using phenol:choloform or the QIAEX desalting protocol. |
| Dephosphorylation of DNA Fragments with Alkaline Phosphatase - http://cshprotocols.cshlp.org/cgi/content/full/2006/2/pdb.prot3863?print=true Dephosphorylation of DNA Fragments with Alkaline Phosphatase. Joseph Sambrook and David W. Russell. This protocol was adapted from Molecular Cloning, ... |
| Protocol: Vector Dephosphorylation Protocol, NEB - http://www.neb.com/nebecomm/products/protocol76.asp Vector Dephosphorylation Protocol. Protocol. Add 1/10 volume of 10X Antarctic Phosphatase Reaction Buffer to 1-5 µg of DNA cut with any restriction ... |
| Protocol for Dephosphorylation of DNA 5'-termini - http://www.fermentas.com/techinfo/modifyingenzymes/protocols/p_dephosph53dna_sap.htm Protocol for Dephosphorylation of DNA 5'-termini (with Shrimp Alkaline Phosphatase (SAP)). Prepare the following reaction mixture: ... |
| IGEM:Harvard/2007/Protocols/Vector Dephosphorylation Protocol ... - http://openwetware.org/wiki/IGEM:Harvard/2007/Protocols/Vector_Dephosphorylation_Protocol Dephosphorylation Protocol ... Buffer to 1 µg of DNA cut with any restriction endonuclease in any buffer. ... |
| DNA Blunting Protocol - http://www.hawaii.edu/microbiology/MO/blunting.htm Aug 10, 2003 ... DNA Blunting Protocol (Revised from Takara Shuzo Co., Ltd.) [back]. I. Description. II. Protocols. A. Blunting Reaction B. Dephosphorylation ... |
