DESIGN AND APPLICATION OF rtPCR METHOD FOR ANALYSIS OF STRAIN-PRODUCER RESIDUAL DNA IN HISTON H1.3 SUBSTANCE

Autor:	A. P. Barannik, E. D. Shibanova, V. I. Shvets, E. V. Kovaleva, Yu. S. Skoblov
Rok vydání:	2015
Předmět:	Ecology Strain (chemistry) Eukaryotic DNA replication Biology Amplicon 16S ribosomal RNA Applied Microbiology and Biotechnology Molecular biology law.invention chemistry.chemical_compound Real-time polymerase chain reaction chemistry law Recombinant DNA Gene DNA Biotechnology
Zdroj:	Biotekhnologiya. :65-75
ISSN:	2500-2341 0234-2758
Popis:	The application of the real-time PCR quantitative method (rtPCR) with specific primers to the 16S RNA E.coli gene fragment for the detection of residual DNA of E. coli BL21(DE3)/ prhH1.3 strain-producer in the APS of the recombinant human histon H1.3 has been suggested. The sample preparation was optimized, a working standard based on the sonicated total DNA of the strain-producer was designed, and an amplicon for the rtPCR was selected. The specifity of the designed primers was evaluated in comparison with eukaryotic DNA. The efficiency of the method in comparison with that using a commercial kit was analyzed. The designed primers to the 16S RNA gene proved to be by 30 times more sensitive than primers of a reference commercial kit.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::d8b51e92a41bf5b611f9d9c4290174ec https://doi.org/10.21519/0234-2758-2015-2-65-75 Zobrazit plný text záznamu