Metatranscriptomics of the heart and kidney tissue of a Costa Rican pygmy rice rat (Oligoryzomys costaricensis)
We performed RNA extraction on frozen tissue using the QIAamp viral RNA minikit according to the manufacturers instructions. Utilizing the Zymo RNA Clean & Concentrator-5 kit, extracted RNA was concentrated and treated with DNAse I utilizing on-column digestion. The resulting RNA was depleted of ribosomal RNA for two hours, converted to cDNA using random hexamers, and i7 and i5 sequencing adaptors added. Finally, individual samples were barcoded and amplified by PCR. All depletion and library preparation steps were conducted using the Zymo-Seq RiboFree Total RNA Kit following the manufacturers recommended protocol.