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Chapters
- 00:02Title
- 00:08Introduction
- 00:48Mosquito Rearing
- 01:31Mosquito gut dissection
- 01:43Metagenomic DNA and RNA isolation
- 02:27Ribosomal RNA depletion
- 02:34PCR amplification of rRNA gene fragments
- 02:48In vitro transcription of rRNA probes
- 03:19rRNA subtraction
- 03:39Removal of rRNA
- 05:07Representative results
- 05:48Conclusion
A ribosomal RNA (rRNA) depletion protocol was developed to enrich messenger RNA (mRNA) for RNA-seq of the mosquito gut metatranscriptome. Sample specific rRNA probes, which were used to remove rRNA via subtraction, were created from the mosquito and its gut microbes. Performance of the protocol can result in the removal of approximately 90-99% of rRNA.
