PMID: 375192Mar 1, 1979Paper

Addition of oligonucleotides to the 5'-terminus of DNA by T4 RNA ligase

Nucleic Acids Research
N P HigginsN R Cozzarelli

Abstract

Bacteriophage T4-induced RNA ligase catalyzes the controlled template-independent addition of RNA to the 5'-phosphoryl end of large DNA molecules. Restriction enzyme-generated fragments of Co1E1 DNA with completely basepaired or cohesive ends and linear single-stranded øX174 viral DNA were all good substrates. DNA molecules from 10 to 6000 nucleotides long were quantitatively joined in an hour to a number of different RNA homopolymers. The most efficient of these was A(pA)5; I(pI)5 and C(pC)5 were also utilized while U(pU)5 was not. The optimum ribohomopolymer length was six nucleotides. Joining of ribohomopolymers between 10 and 20 nucleotides long occurred at approximately 1/2 the maximal rate and a trimer was the shortest substrate. Thus RNA ligase provides a method for generating extensions of predetermined length and base composition at the 5'-end of large DNA molecules that complements the available procedures for extending the 3'-hydroxyl terminus of DNA.

References

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Citations

Jan 1, 1993·Bio Systems·A B Chetverin, F R Kramer
Jul 17, 2008·Nucleic Acids Research·Omid R FaridaniLiam Good
Sep 1, 1982·Analytical Biochemistry·H Mei-haoH Hui-fen
Oct 1, 1986·Analytical Biochemistry·D C TessierT Vernet

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