title: Bacterial 16S rRNA gene library from pot experiment with clean sand and willows, growth time 60 days
design: Amplicon libraries for the bacterial 16S rRNA gene variable regions 18 (bV18?A) were generated using up to 25?ng DNA as PCR template. PCR reaction (50?L) contained 0.2mM dNTP mix, 0.01 units of Platinum SuperFi DNA Polymerase, and 500 nM primers containing custom 24nt barcodes followed by [8F] AGRGTTYGATYMTGGCTCAG and [1391R] GACGGGCGGTGWGTRCA, using a program of 98C 3min; 25cycles of 98C 30s, 62C 20s, 72C 2min; and 72C 5min, after which amplicons were purified with CleanNGS SPRI beads (3:5 ratio), eluted in 25 L nuclease?free water, quantified by Qubit, assessed by Tapestation 2200 and D1000/High sensitivity D1000 screentap for size and purity, and used to prepare sequencing libraries with the SQK?LSK114 kit with 500?ng input and SPRI?based clean?ups