Author: Girardi, Erika; Chane-Woon-Ming, Béatrice; Messmer, Mélanie; Kaukinen, Pasi; Pfeffer, Sébastien
Title: Identification of RNase L-Dependent, 3'-End-Modified, Viral Small RNAs in Sindbis Virus-Infected Mammalian Cells Document date: 2013_11_19
ID: v6uc0ijw_29
Snippet: 3=-end modification is a hallmark of SvsRNAs. Taken together, our observations indicate that the SINV genomic RNA is modified, and are therefore suggestive of a potential modification of SvsRNAs. We then decided to investigate to what extent reads mapping to the SINV genome were modified at the last nucleotide position. Small RNA libraries were prepared from noninfected or SINV-infected HEK293 cells using total RNA treated (or not) with NaIO 4 . .....
Document: 3=-end modification is a hallmark of SvsRNAs. Taken together, our observations indicate that the SINV genomic RNA is modified, and are therefore suggestive of a potential modification of SvsRNAs. We then decided to investigate to what extent reads mapping to the SINV genome were modified at the last nucleotide position. Small RNA libraries were prepared from noninfected or SINV-infected HEK293 cells using total RNA treated (or not) with NaIO 4 . This treatment specifically impairs the 3= adapter ligation to RNA species with unmodified 3= ends (2= and 3= OH), while RNAs bearing a modification at one of these positions are resistant to oxidation, and can therefore be ligated. Overnight ligation of the 3= adapter at 16°C was performed in order to enhance the cloning of 2=-O modified small RNAs (39) .
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