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排序方式: 共有120条查询结果,搜索用时 955 毫秒
61.
Lapa S. A. Pavlov A. S. Kuznetsova V. E. Shershov V. E. Spitsyn M. A. Guseinov T. O. Radko S. P. Zasedatelev A. S. Lisitsa A. V. Chudinov A. V. 《Molecular Biology》2019,53(3):460-469
Molecular Biology - The effects of modified deoxyuridine triphosphates (mod-dUTPs) with different substituents at the C5 position of the pyrimidine cycle on the kinetics of PCR with Taq and Vent... 相似文献
62.
Fedorova OE Liubchenko LN Paiadini IuG Kazubskaia TP Amosenko FA Gar'kavtseva RF Zasedatelev AS Nasedkina TV 《Molekuliarnaia biologiia》2007,41(1):37-42
Ovarian cancer (OC) is one of the leading cause of cancer death in women. Inherited BRCA1 and BRCA2 mutations strikingly increase OC risk (with lifetime risk estimates ranging at 10-60%). Mutation 1100delC in CHEK2 gene was shown to be associated with breast cancer in women carrying this mutation. Knowledge of the nature and frequency of population-specific mutations in these genes is a critical step in the development of simple and inexpensive diagnostic approaches to DNA analysis. The frequencies of 185delAG, 300T>G, 4153delA, 4158A>G, 5382insC mutations in BRCA1 gene, 695insT and 6174delT mutations in BRCA2 gene and 1100delC mutation in CHEK2 gene were analyzed using biochips in Russian OC patients. We studied 68 women who received a diagnosis of epithelial OC and 19 women with primary multiple tumors involving the ovaries. The 185delAG, 300T>G, 4153delA and 5382insC in BRCA1 gene were identified. The most prevailing mutation was 5382insC in BRCA1 gene (87.5% of all BRCA1 mutations OC patients, 50.0% in patients with primary multiple tumors involving the ovaries). No mutations in BRCA2 and CHEK2 genes were detected. 相似文献
63.
The article reviews the last period of A. D. Mirzabekov's scientific career. During this time, gel-based biochips were invented, studied, and introduced to practical applications in his laboratory. This work began at the early stages of the "Human Genome" project and is continuing today, including recent development of diagnostic oligonucleotide and protein biochips. This research is discussed in the context of the worldwide development of microarray technologies. 相似文献
64.
Fesenko D. O. Ivanovsky I. D. Ivanov P. L. Zemskova E. Yu. Agapitova A. S. Polyakov S. A. Fesenko O. E. Filippova M. A. Zasedatelev A. S. 《Molecular Biology》2022,56(5):780-799
Molecular Biology - This paper presents a method for genotyping a panel of 60 single nucleotide polymorphisms (SNPs) using single-stage PCR followed by hybridization on a hydrogel biochip. The pool... 相似文献
65.
E. E. Belobritskaya M. V. Neunylova V. A. Vasiliskov V. D. Rumyantseva A. V. Chudinov A. S. Zasedatelev 《Russian Journal of Bioorganic Chemistry》2007,33(6):617-619
A number of boradiazaindacene dyes containing a carboxyl group separated from a fluorophore by two methylene units were synthesized. The compounds have narrow spectral bands with absorption maxima at 480–530 nm and fluorescence maxima at 500–550 nm. Succinimide esters of these compounds and the corresponding fluorescent-labeled oligonucleotides were also prepared. Boradiazaindacene dyes can be used as fluorescent labels for oligonucleotides for analysis of melting curves of duplexes on microchips either by themselves or in combination with Texas Red. They can also be applied for labeling primers for polymerase chain reaction. 相似文献
66.
Sorokin NV Chechetkin VR Livshits MA Pan'kov SV Donnikov MY Gryadunov DA Lapa SA Zasedatelev AS 《Journal of biomolecular structure & dynamics》2005,22(6):725-734
The efficiency of discrimination between perfect and mismatched duplexes during hybridization on microchips depends on the concentrations of target DNA in solution and immobilized probes, buffer composition, and temperature of hybridization and is determined by both thermodynamic relationships and hybridization kinetics. In this work, optimal conditions of discrimination were studied using hybridization of fluorescently labeled target DNA with custom-made gel-based oligonucleotide microchips. The higher the concentration of immobilized probes and the higher the association constant, the higher the concentration of the formed duplexes and the stronger the corresponding fluorescence signal, but, simultaneously, the longer the time needed to reach equilibrium. Since mismatched duplexes hybridize faster than their perfect counterparts, perfect-to-mismatch signal ratio is lower in transient regime, and short hybridization times may hamper the detection of mutations. The saturation time can be shortened by decreasing the probe concentration or augmenting the gel porosity. This improves the detection of mutations in transient regime. It is shown that the decrease in the initial concentration of oligonucleotide probes by an order of magnitude causes only 1.5-2.5-fold decrease of fluorescence signals after hybridization of perfect duplexes for 3-12 h. At the same time, these conditions improve the discrimination between perfect and mismatched duplexes more than two-fold. A similar improvement may be obtained using an optimized dissociation procedure. 相似文献
67.
Kolchinskiĭ AM Griadunov DA Lysov IuP Mikhaĭlovich VM Nasedkina TV Turygin AIu Rubina AIu Barskiĭ VE Zasedatelev AS 《Molekuliarnaia biologiia》2004,38(1):5-16
The review describes the history of creation and development of the microchip technology and its role in the human genome project in Russia. The emphasis is placed on the three-dimensional gel-based microchips developed at the Center of Biological Microchips headed by A.D. Mirzabekov since 1988. The gel-based chips of the last generation, IMAGE chips (Immobilized Micro Array of Gel Elements), have a number of advantages over the previous versions. The microchips are manufactured by photo-initiated copolymerization of gel components and immobilized molecules (DNA, proteins, and ligands). This ensures an even distribution of the immobilized probe throughout the microchip gel element with a high yield (about 50% for oligonucleotides). The use of methacrylamide as a main component of the polymerization mixture resulted in a substantial increase of gel porosity without affecting its mechanical strength and stability, which allowed one to work with the DNA fragments of up to 500 nt in length, as well as with rather large protein molecules. At present, the gel-based microchips are widely applied to address different problems. The generic microchips containing a complete set of possible hexanucleotides are used to reveal the DNA motifs binding with different proteins and to study the DNA-protein interactions. The oligonucleotide microchips are a cheap and reliable tool of diagnostics designed for mass application. Biochips have been developed for identification of the tuberculosis pathogen and its antibiotic-resistant forms; for diagnostics of orthopoxviruses, including the smallpox virus; for diagnostics of the anthrax pathogen; and for identification of chromosomal rearrangements in leukemia patients. The protein microchips can be adapted for further use in proteomics. Bacterial and yeast cells were also immobilized in the gel, maintaining their viability, which open a wide potential for creation biosensors on the basis of microchips. 相似文献
68.
T.?O.?Guseinov V.?E.?Kuznetsova V.?E.?Shershov M.?A.?Spitsyn S.?A.?Lapa A.?S.?Zasedatelev A.?V.?ChudinovEmail author 《Russian Journal of Bioorganic Chemistry》2018,44(2):252-255
Herein, we present a new synthetic route to fluorescently-labeled nucleoside triphosphates via Sonogashira cross-coupling of iodinated deoxycytidine monophosphate and cyanine dye with a terminal alkyne group. 相似文献
69.
D.?O.?Fesenko T.?O.?Guseinov S.?A.?Lapa V.?E.?Kuznetsova V.?E.?Shershov M.?A.?Spitsyn T.?V.?Nasedkina A.?S.?Zasedatelev A.?V.?ChudinovEmail author 《Molecular Biology》2018,52(3):458-466
The efficiency of the incorporation of fluorescently labeled derivatives of 2'-deoxycytidine in DNA synthesized de novo has been studied using PCR with Taq and Tth polymerases of family A and Vent (exo–) and Deep Vent (exo–) polymerases of family B. Four derivatives of 5'-triphosphate-2'-deoxycytidine (dCTP) have different chemical structures of the indodicarbocyanine dye and Cy5 analogue attached to position 5 of cytosine. The kinetics of the accumulation of the PCR products and the intensity of the fluorescent signals in the hybridization analysis with immobilized DNA probes depend on the modification of the fluorescently labeled dCTP counterpart, its concentration, and the type of DNA polymerase. All labeled triphosphates showed some inhibitory effects on PCR. The best balance between the efficiency of incorporating labeled cytidine derivatives and the negative effect on the PCR kinetics has been shown in the case of Hot Taq polymerase in combination with the Cy5-dCTP analogue, which contains containing electrically neutral chromophore, the axis of which is a continuation of the linker between the chromophore and the pyrimidine base. 相似文献
70.
Anna V. Kudryavtseva Maria S. Fedorova Alex Zhavoronkov Alexey A. Moskalev Alexander S. Zasedatelev Alexey A. Dmitriev Asiya F. Sadritdinova Irina Y. Karpova Kirill M. Nyushko Dmitry V. Kalinin Nadezhda N. Volchenko Nataliya V. Melnikova Kseniya M. Klimina Dmitry V. Sidorov Anatoly Y. Popov Tatiana V. Nasedkina Andrey D. Kaprin Boris Y. Alekseev George S. Krasnov Anastasiya V. Snezhkina 《BMC genetics》2016,17(3):156