By Leen Vijgen PhD, Elien Moës, Els Keyaerts PhD, Sandra Li, Marc Van Ranst MD, PhD (auth.), Dave Cavanagh (eds.)
In 2003, the note "coronavirus" unfold around the globe, a bit extra than the virus that sparked the panic. In SARS- and different Coronaviruses: Laboratory Protocols, professional researchers study those devastating viruses via special laboratory protocols. Chapters care for such topics as detection and discovery of coronaviruses by means of nucleic acid and antibody-based ways, virus isolation, propagation and titration, virus purification, constitution research via electron cryomicroscopy, expression and crystallization of viral proteins, elevating antibodies opposed to viral proteins, manipulation of the coronavirus genome, and outlines of ways to enquire elements of the mobilephone floor for coronavirus receptors. Composed within the hugely winning Methods in Molecular Biology™ sequence structure, every one bankruptcy encompasses a short advent, step by step tools, a listing of useful fabrics, and a Notes part which stocks pointers on troubleshooting and averting recognized pitfalls.
Comprehensive and state-of-the-art, SARS- and different Coronaviruses: Laboratory Protocols serves as a fantastic advisor for all virologists and particularly for these operating with coronaviruses.
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Extra resources for SARS- and Other Coronaviruses: Laboratory Protocols
Sample text
In our experience, better amplification of RNA products is obtained when mineral oil is added to the reaction, regardless of whether or not a thermocycler, with or without, a hot lid is used. 12. Even the conserved, 3 -terminal part of the 3 UTR of group 3 avian coronaviruses exhibits some variation. It is likely that oligonucleotides UTR11– and UTR41+ will require modification to increase their universality for group 3 coronaviruses. For example, we modified our initial versions of these primers [UTR1and UTR4+ (12)] to the current UTR11– and UTR41+ as more IBV sequences became available.
2005) The structure of a rigorously conserved RNA element within the SARS virus genome PLoS Biol. 3, E5. 3. Jonassen, C. , Larsen, I. , Follestad, A. and Lillehaug, A. (2005) Molecular identification and characterization of novel coronaviruses infecting graylag geese (Anser anser), feral pigeons (Columbia livia) and mallards (Anas platyrhynchos) J. Gen. Virol. 86, 1597–1607. 4. Ksiazek, T. , Goldsmith, C. , Zaki, S. , Comer, J. , Rollin, P. , Dowell, S. , Ling, A. , Humphrey, C. , Shieh, W. , Paddock, C.
37 27586 27342 14184 13953 IBV 5’ positiona 27607 27364 14204 13976 IBV 3’ positiona 56 60 53 52 Tm (◦ C) 3’ UTR 3’ UTR gene 1 gene 1 Region amplified I, T, Ph I, T, Ph I, T, Ph, Pa I, T, Ph, Pa Avian CoV amplifiedb (2) (2) (1) (1) Reference b Relative to the genome of the Beaudette strain of IBV (Accession number AJ311317). The two primer combinations were tested using RNA extracted from IBV (I), TCoV, (T), pheasant coronavirus (PhCoV), and parrot coronavirus (PaCoV). a 22 UTR11– – + 23 gct cta act cta tac tag cct a – 20 Sense + Length 23 Sequence act ca(ag) (at)t(ag) aat (ct) t(agct) aaa ta(ct) gc 4 Bm tca ca(ct) tt(at) gga ta(ag) tcc ca UTR41+ atg tct atc gcc agg gaa atg tc 2 Bp Primer name Table 1 Details of the Oligonucleotide Primers Described in this Chapter (see Note 12) 38 Culver et al.