16S amplicon sequencing for C prolifera
The DNA and cDNA samples were subsequently used in a two-stage PCR protocol (Naqib et al., 2018) to amplify the V5-V7 region of the 16S rRNA gene. The first stage PCR was done with CS1_799F - CS2-1193R chloroplast-avoiding primers (Beckers et al., 2016) using 1 ng of DNA per 10 L reaction. Subsequently, a second-stage PCR was done with the barcoded primers (Naqib et al., 2018). A mock community (MSA-1002, ATCC) was used as a positive control, and molecular biology-grade water was used as a negative control.