title: 16S amplicon sequencing of feces from C57BL/6J mice
design: Primers 515F and 806R were used to amplify the V4 domain of the 16S rRNA. Each sample was barcoded with a unique forward and reverse barcode combination. PCR contained 1 Unit Kapa2G Robust Hot Start Polymerase, 1.5mM MgCl2, 10 mol of each primer, 10mM dNTP s, and 1ul of DNA. PCR conditions were: an initial incubation at 95 C for 2 min, followed by 30 cycles of 95 C for 20 s, 50 C for 20 s, 72 C for 20 s and a final extension of 72 C for 3 min. The final product was quantified on the Qubit instrument using the Qubit High Sensitivity DNA kit and individual amplicon libraries were pooled.
library_name: B6J.rep2