Lb. fermentum KUB-D18 genome
Throughout the whole process of sequencing from the DNA sample to the final data, each step, including sample test, library preparation and sequencing procedures, influences the quality of data production, while the data quality further impacts on the analysis results directly. To guarantee the accuracy and reliability of the sequencing data, Novogene utilizes stringent quality control (QC) procedures and strictly adheres to the high standard at each step from source. The genomic DNA of each sample was randomly sheared into short fragments of about 350 bp respectively. The obtained fragments were subjected to library construction using the NEBNext DNA Library Prep Kit, following the instructions strictly. Briefly, as followed by end repairing, dA-tailing, and further ligation with NEBNext adapter, the required fragments (in 300-500 bp size) were PCR enriched by P5 and indexed P7 oligos. After purification and subsequent quality check, the library is ready for sequencing. To check the prepared DNA libraries, Qubit 2.0 fluorometer was firstly used to determine the concentration of the library. After dilution to 1 ng/ul, the Agilent 2100 bioanalyzer was used to assess the insert size. And finally the quantitative real-time PCR (qPCR) was performed to detect the effective concentration of each library. If the library with appropriate insert size has an effective concentration (more than 2 nM), the constructed libraries are qualified and ready for Illumina high-throughput sequencing. The qualified DNA libraries were pooled according to their effective concentration as well as the expected data production. Pair-end sequencing were performed on Illumina sequencing platform, with the read length of PE150 bp at each end.