| Popis: |
Extracting bulk-RNA from biospecimens is a time-consuming process. Column-based extractions performed by hand must be done in batches and the number of samples that can be extracted per batch is dependent on a single person's abilities. Hence, extraction hundreds of samples can take weeks. With every round of extractions, reagents are susceptible to differing amounts of contamination and technical error, resulting in batch effects. Microbial RNA input for metagenomic next-generation sequencing is often found in very low quantities; therefore, contamination introduced during extraction may influence sequencing results and depict an inaccurate reflection of microbes present in the original sample. Here, we outline a high-throughput, semi-automatic method to extract RNA using a Agilent Bravo liquid handler. This protocol utilizes a magnetic bead-based approach to extracting RNA and is performed in 96-well deep-well plates. All reagents are plated into 96-well deep-well plates before the start of the Bravo protocol. The liquid handler is programmed to aliquot, mix, and aspirate reagents from different decks of plates. The user is only responsible for putting these plates on the correct decks. With this approach, we can extract a full plate of 96 samples within a day and reduce the susceptibility to contamination from user error. |
