Microbial RNAtag-seq

Input Requirement(s)
500ng of total RNA at a concentration of 25ng/ul

Batch Size: 32 samples or 96 samples

Our RNAtag-seq process (Shishkin et. al., 2015) provides high complexity transcriptome sequencing libraries without the requirement for a polyA selection step. The approach is strand specific and yields quantitative data suitable for transcriptome profiling. This process was originally designed for microbial transcriptome profiling as it does not rely on oligo(dT) priming, rather the first step involves RNA ligation of a primer to the transcript. While best results are typically achieved with high quality RNA (RIN >7), the approach is effective for low quality or degraded RNA, with RIN scores as low as 2. Importantly, a unique molecular barcode is attached to individual samples at an early stage in the workflow, enabling pooling early in the process to increase efficiency. RNAtag-seq is frequently used for expression analysis studies including: human microbial commensals and pathogens, intracellular microbial pathogens, metagenomic/metatranscriptomic populations, analysis of antibiotic response signatures, regulatory circuit analysis in support of synthetic biology work and annotation of genomes.