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Methods Mol Biol. 2014;1093:209-24. doi: 10.1007/978-1-62703-694-8_17.

Analysis of Piwi-loaded small RNAs in Tetrahymena.

Author information

1
Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria.

Erratum in

  • Methods Mol Biol. 2014;1093:E1-2.

Abstract

Scan RNAs (scnRNAs) are developmentally regulated siRNAs of ~26-32 nucleotides in length that are involved in programmed DNA elimination in Tetrahymena. scnRNAs are loaded onto the Piwi-related protein Twi1p and 2'-O-methylated at their 3' termini. We describe two alternative strategies for analyzing the Twi1p-loaded scnRNAs: preparation of loaded scnRNAs by immuno-purification of the Twi1p-scnRNA complex and exclusion of non-methylated scnRNAs during cDNA library construction using periodate oxidation.

PMID:
24178568
DOI:
10.1007/978-1-62703-694-8_17
[Indexed for MEDLINE]

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