sequences of bacteria
After thawing at 25C, 10 g tea leaves were transferred in polypropylene test tubes containing 40 mL sterile water and orbit-shaked at 150rpm for 1 h at room temperature. The suspensions were separated using 0.22 m vacuum filtration units (EMD Millipore, Billerica, MA) and the sediment filtered were used as samples for the extraction of microbial DNA using HiPure Soil DNA Kits (Magen, Guangzhou, China). All amplifications were performed in triplicate 50 L reactions, including: DNA template (50 ng) 1.0 L, dNTPs (2.5 mmol/L) 1.5 L, each primer (10 mol/L) 1.5 L, 5 Q5 reaction buffer (New England Biolabs) 10 L, 5 Q5 High GC Enhancer 10 L, Q5 High-Fidelity DNA Polymerase (New England Biolabs) 0.3 L, and ddH2O 24.2 L. Thermal cycler conditions consisted of an initial denaturation at 95C for 5 min, then 30 cycles of 95C for 1 min, 60C for 1 min, 72C for 1 min, and a final extension at 72C for 7 min. It was purified using AxyPrep DNA Gel Extraction Kit from Axygen Biosciences, Union City, US, and quantified using Life Technologies' ABI StepOnePlus Real-Time PCR System. The sequence libraries were pooled into equimolar quantities and paired-end sequenced (PE250) on the Illumina platform (San Diego, US).