GSM9462328: RNA_pool_2; Escherichia coli DH5[alpha]; RNA-Seq
Cells were harvested from the pooled cultures at a defined time point. Total RNA was extracted using the RNAprep Pure Cell/Bacteria Kit according to the manufacturer's instructions. In parallel, plasmid DNA was extracted from the same cultures to assess construct abundance in the library. For RNA-derived samples, total RNA was reverse transcribed into cDNA using the HiScript IV 1st Strand cDNA Synthesis Kit. Barcode-containing regions were selectively amplified by PCR using the primers Test-bar-F/R. For DNA-derived samples, barcode regions were directly amplified from plasmid DNA using the same primer pair. Sequencing libraries were constructed from the amplified products following standard Illumina protocols.