wetland4
Total RNA was extracted from sediment samples collected from wetlands using the RNeasy Micro Kit (Qiagen). RNA quality and concentration were assessed using a Nanodrop spectrophotometer and agarose gel electrophoresis. Ribosomal RNA was depleted using the QIAseq FastSelect 5S/16S/23S kit with the KAPA RNA HyperPrep Kit. The resulting RNA was purified, reverse-transcribed into cDNA, and amplified. Libraries were purified using KAPA Pure Beads and quantified with a Qubit 3.0 Fluorometer. The prepared cDNA libraries were subjected to de novo metatranscriptomic sequencing to profile microbial community gene expression in sediment samples.