Počet záznamů: 1

In situ reverse transcription: the magic of strength and anonymity

  1. 1.
    SYSNO ASEP0345708
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    NázevIn situ reverse transcription: the magic of strength and anonymity
    Tvůrce(i)Ligasová, Anna (UEM-P) - Koberna, Karel (UEM-P)
    Rozsah strane167 - e178
    Zdroj.dok.Nucleic Acids Research - ISSN 0305-1048
    Poč.str.12 s.
    Jazyk dok.eng - angličtina
    Země vyd.GB - Velká Británie
    Klíč. slovanucleic acids ; DNA-RNA
    Vědní obor RIVEA - Morfologické obory a cytologie
    CEPGA204/09/0973 GA ČR - Grantová agentura ČR
    KJB500390701 GA AV ČR - Akademie věd
    KAN200520801 GA AV ČR - Akademie věd
    CEZAV0Z50390512 - UEM-P (2005-2011)
    UTISI000281720500007
    DOI10.1093/nar/gkq619
    AnotaceIn this study, we describe an approach that enables a highly specific, effective and fast detection of polyadenylated RNA sequences in situ at the light and electron microscopy levels. The method developed is based on the incorporation of 5-bromo-2’-deoxyuridine into the growing cDNAstrand by means of the reverse transcriptase. We have shown that unlike the previously used deoxyuridine tagged with biotin or digoxigenin,5-bromo-2‘-deoxyuridine is ‘invisible’ in the DNA–DNA duplex but easily detectable in theDNA–RNA duplex. We have also shown that the replacement of deoxythymidine by 5-bromo-2‘-deoxyuridine considerably stabilizes the growing DNA–RNA duplex, thus enabling the one-step detection of polyadenylated RNA in structurally well-preserved cells.
    PracovištěÚstav experimentální medicíny
    KontaktLenka Koželská, lenka.kozelska@biomed.cas.cz, Tel.: 241 062 218, 296 442 218
    Rok sběru2011