Culturome of leaf microbiome: epiphytes
First and round PCR reaction was done under the following conditions: 2 min at 94 C; 32 cycles of denaturation at 94 C for 30 s, annealing at 55 C for 30 s, and extension at 72 C for 60 s; final step 5 min at 72 C. The 30 L PCR mixture contained 3 L of takara buffer, 0.15 LHS taq, 20.85L Qiagen water, 2.4 L dNTPs, 0.6 L of each primer (10 M) and 3 L template DNA. PCR products were collected and purified using Wizard SV Gel and PCR Clean-up System and Agencourt AMPure XP beads.  Library  sequencing was conducted using the Illumina Novaseq 6000 Sequencer (Illumina, Inc., CA, USA).