Deep hyper saline anoxic basin sediments
After confirming the absence of DNA, the Ovation Single Cell RNA-seq kit         (Nugen, San Carlos, CA) was used to create barcoded cDNA libraries for sequencing. A         modified step was followed in which the First Strand Primer Mix, A1 ver 15 (S01882) was         replaced with the First Strand Primer Mix, A1 ver 13 (S01858) to avoid biases against         prokaryotic transcripts. The libraries were sequenced at DNA Sequencing & Genotyping         Center, University of Delaware using Illumina HiSeq platform and paired-end 150 run. The         five libraries were pooled in 3 lanes. The forward and reverse reads were filtered using         Trimmomatic. The length of the trimmed sequences was set to be at least 50 nt. Surviving         trimmed reads were used for assembly with Trinity release r20140717.