Tn-Seq of GBS: calprotectin
Briefly, genomic DNA was sheared to approximately 340 bp fragments and processed through the Ovation Ultralow V2 DNA-Seq library preparation kit (Tecan) and 9ng of each library was used as template to enrich by PCR (16 cycles) for the transposon insertions using Krmit-specific (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCGGGGACTTATCATCCAACC) and Illumina P7 primers. The enriched PCR products were diluted 1:100, and 20L used as template for an indexing PCR (9 cycles) using the TruSeq P5 indexing and P7 primers. Sequencing was performed using Illumina NovaSeq 6000 in 150 base paired-end format.