Please use this identifier to cite or link to this item:
http://hdl.handle.net/10261/37787
Share/Export:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Title: | Enzymatic synthesis of structure-free DNA with pseudo-complementary properties |
Authors: | Lahoud, Georges; Timoshchuk, Víctor; Lebedev, Alexandre; Vega, Miguel de CSIC ORCID ; Salas, Margarita CSIC ORCID ; Aarar, Khalil; Hou, Ya-Ming; Gamper, Howard | Issue Date: | 29-Apr-2008 | Publisher: | Oxford University Press | Citation: | Nucleic Acids Research 36(10): 3409–341(2008) | Abstract: | Long single-stranded DNAs and RNAs possess considerable secondary structure under conditions that support stable hybrid formation with oligonucleotides. Consequently, different oligomeric probes can hybridize to the same target with efficiencies that vary by several orders of magnitude. The ability to enzymatically generate structure-free singlestranded copies of any nucleic acid without impairing Watson–Crick base pairing to short probes would eliminate this problem and significantly improve the performance of many oligonucleotide-based applications. Synthetic nucleic acids that exhibit these properties are defined as pseudo-complementary. Previously, we described a pseudo-complementary A-T couple consisting of 2-aminoadenine (nA) and 2-thiothymine (sT) bases. The nA-sT couple is a mismatch even though nA-T and A-sT are stable base pairs. Here we show that 7-alkyl-7-deazaguanine and N 4 -alkylcytosine (where alkyl = methyl or ethyl) can be used in conjunction with nA and sT to render DNA largely structure-free and pseudo-complementary. The deoxynucleoside triphosphates (dNTPs) of these bases are incorporated into DNA by selected mesophilic and thermophilic DNA polymerases and the resulting primer extension products hybridize with good specificity and stability to oligonucleotide probes composed of the standard bases. Further optimization and characterization of the synthesis and properties of pseudo-complementary DNA should lead to an ideal target for use with oligonucleotide probes that are <25 nt in length | Publisher version (URL): | http://dx.doi.org/10.1093/nar/gkn209 | URI: | http://hdl.handle.net/10261/37787 | DOI: | 10.1093/nar/gkn20 | ISSN: | 0305-1048 |
Appears in Collections: | (CBM) Artículos |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Nucl. Acids Res.-2008-Lahoud-3409-19.pdf | 3,67 MB | Adobe PDF | View/Open |
CORE Recommender
Page view(s)
269
checked on Apr 19, 2024
Download(s)
135
checked on Apr 19, 2024
Google ScholarTM
Check
Altmetric
Altmetric
WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.