دورية أكاديمية
[Preparation of Modified Combinatorial DNA Libraries via Emulsion PCR with Subsequent Strand Separation].
| العنوان: | [Preparation of Modified Combinatorial DNA Libraries via Emulsion PCR with Subsequent Strand Separation]. |
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| المؤلفون: | Lapa SA; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.; lapa@biochip.ru., Romashova KS; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia., Spitsyn MA; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.; IBMC-EcoBioPharm Ltd., Moscow, 119121 Russia., Shershov VE; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia., Kuznetsova VE; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia., Guseinov TO; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.; IBMC-EcoBioPharm Ltd., Moscow, 119121 Russia., Zasedateleva OA; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia., Radko SP; IBMC-EcoBioPharm Ltd., Moscow, 119121 Russia.; Orekhovich Institute of Biomedical Chemistry, Russian Academy of Sciences, Moscow, 119121 Russia., Timofeev EN; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia., Lisitsa AV; Orekhovich Institute of Biomedical Chemistry, Russian Academy of Sciences, Moscow, 119121 Russia., Chudinov AV; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.; IBMC-EcoBioPharm Ltd., Moscow, 119121 Russia. |
| المصدر: | Molekuliarnaia biologiia [Mol Biol (Mosk)] 2018 Nov-Dec; Vol. 52 (6), pp. 984-996. |
| نوع المنشور: | Journal Article |
| اللغة: | Russian |
| بيانات الدورية: | Publisher: Izdatelstvo Nauka Country of Publication: Russia (Federation) NLM ID: 0105454 Publication Model: Print Cited Medium: Print ISSN: 0026-8984 (Print) Linking ISSN: 00268984 NLM ISO Abbreviation: Mol Biol (Mosk) Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Moskva : Izdatelstvo Nauka |
| مواضيع طبية MeSH: | Aptamers, Nucleotide* , Gene Library* , Polymerase Chain Reaction* , SELEX Aptamer Technique*, DNA/*isolation & purification |
| مستخلص: | A modification of the enzymatic method for the preparation of combinatorial random DNA libraries, which combines amplification in isolated microvolumes with the simultaneous incorporation of modified nucleotides and subsequent separation of DNA strands, was developed. Deoxyuridine triphosphate with hydrophobic substituents such as structural analogues of amino acid side chains in the C5 position of the pyrimidine ring was used to introduce modifications into DNA. To prevent competitive amplification, which reduces the representativeness of combinatorial libraries, PCR in inverse emulsion was used. The separation of the strands of PCR products was carried out. There were six single-stranded DNA libraries with complete substitution of deoxythymidine via modified analogues with various functional groups. These DNA libraries are suitable for generating aptamers to protein targets through additional hydrophobic interactions from the introductions of appropriate modifications, and are completely compatible with the SELEX aptamer selection methodology. |
| فهرسة مساهمة: | Keywords: 2'-deoxyuridine-5'-triphosphate; DNA polymerases; PCR in inverse emulsion; combinatorial DNA libraries; modified aptamers; modified nucleotides |
| المشرفين على المادة: | 0 (Aptamers, Nucleotide) 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20190112 Date Completed: 20190620 Latest Revision: 20190620 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1134/S0026898418060113 |
| PMID: | 30633241 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0026-8984 |
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| DOI: | 10.1134/S0026898418060113 |