Abstract
At the current rate of technological progress, high-throughput sequencing of nucleic acids has become a commodity. These techniques are perfectly suitable for viral small RNAs sequencing and contribute to the understanding of many aspects of virus biology in the context of host-pathogen interaction. However, the generation of high quality data is still an issue and the preparation of small RNAs libraries that accurately reflect the viral siRNAs in the sample remains a challenge. In this chapter we describe how to clone and sequence libraries of viral small RNAs from infected insect samples (mosquito, drosophilidae, insect-derived cell lines).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Cloning, Molecular / methods*
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Computational Biology
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DNA Primers / genetics
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DNA Primers / metabolism
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Deoxyribonucleases, Type II Site-Specific / metabolism
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Isotope Labeling
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Oligonucleotides / chemistry
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Oligonucleotides / genetics
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Oligonucleotides / isolation & purification
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Oligonucleotides / metabolism
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Polymerase Chain Reaction
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RNA, Small Interfering / chemistry
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RNA, Small Interfering / genetics*
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RNA, Small Interfering / isolation & purification
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RNA, Small Interfering / metabolism
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RNA, Viral / chemistry
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RNA, Viral / genetics*
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RNA, Viral / isolation & purification
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RNA, Viral / metabolism
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Reverse Transcription
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Sequence Analysis, RNA / methods*
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Viruses / genetics*
Substances
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DNA Primers
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Oligonucleotides
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RNA, Small Interfering
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RNA, Viral
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endodeoxyribonuclease PmeI
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Deoxyribonucleases, Type II Site-Specific