Preparation of short cytosine-modified oligonucleotides by nicking enzyme amplification reaction.
A method for enzymatic production of short (10-20 nt) cytosine-modified oligonucleotides was developed by nicking enzyme amplification reaction using Vent(exo-) polymerase, Nt.BstNBI nicking endonuclease and 5-substituted dCTP derivatives. The methodology including isolation was scaled up to nanomolar amounts and was proved to be suitable for production of diverse base-modified short single-stranded oligonucleotides (inaccessible by other enzymatic methods) that are of potential interest as labelled primers or functionalized aptamers.
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610 Prague 6, Czech Republic.
Petra
P
Ménová
Michal
M
Hocek
D053904:DNA Breaks, Single-Stranded; D017931:DNA Primers / Q000378:metabolism; D003845:Deoxycytosine Nucleotides / Q000737:chemistry*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D009841:Oligonucleotides / Q000096:biosynthesis*
D017931:DNA Primers; D003845:Deoxycytosine Nucleotides; D009841:Oligonucleotides; C024107:2'-deoxycytidine 5'-triphosphate; C430823:endodeoxyribonuclease BstNBI; D015252:Deoxyribonucleases, Type II Site-Specific