Published in
Humana Press, Methods in Molecular Biology, p. 281-293, 2011
DOI: 10.1007/978-1-61779-046-1_18
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Immunoprecipitation of piRNPs and Directional, Next Generation Sequencing of piRNAs
,
Anastassios Vourekas,
Eugene Khandros,
Zissimos Mourelatos
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Abstract
Piwi interacting RNAs (piRNAs) are small ( approximately 25 to approximately 30 nucleotide) and are expressed in the germline. piRNAs bind to the Piwi subclade of Argonaute proteins and form the core ribonucleoproteins (RNPs) of piRNPs. We describe a method for the massive identification of piRNAs from immunopurified piRNPs. This strategy may also be used for immunopurification and directional sequencing of RNAs from other RNPs that contain small RNAs.