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The DNA was extracted from 200 mg cecal samples using the CTAB method. Polymerase chain reaction (PCR) amplification of full length 16S rRNA genes (V1-V9) was performed with barcoded primers.After mixing the PCR products in equidensity ratios, the mixture was purified with a QIAquick@ Gel Extraction Kit (Qiagen, Hilden, Germany). Sequencing libraries were generated using a SMRTbell Template Prep Kit (PacBio, Menlo Park, CA, USA) following the manufacturer's recommendations, and the library quality was assessed on a fluorometer (Qubit 2.0, Thermo-Fisher Scientific, Waltham, MA, USA). The library was sent for sequencing on the PacBio Sequel platform (PacBio). |
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