modified-iPSC.R1.fq.gz
This library prep set is optimized to convert 5-400 ng genomic DNA into a customized library and uses advanced Adapter Ligation technology and High-fidelity PCR Enzymes to significantly increase library yield and conversion rate. Library preparation was performed with MGIEasy FS DNA library prep kit (MGI) according to the manufacturer's instructions. Briefly, After Size-selection of fragmented gDNA using AMPure XP magnetic beads, the fragmented gDNA was end-repaired and a-tailed at 37 C for 30 min, and 65 C for 15 min. Indexing adapter was ligated to the ends of the DNA fragments at 23 C for 30 min. After cleanup of adapter-ligated DNA, PCR was performed to enrich those DNA fragments that have adapter molecules. Thermocycler conditions were as follows: 95 C for 3 min, 8 cycles of 98 C for 20 s, 60 C for 15 s, and 72 C for 30 s, with a final extension at 72 C for 10 min. The double stranded library is quantified using QauntiFluor ONE dsDNA System (Promega. The library is circularized at 37 C for 30 min, and then digested at 37 C for 30 min, followed by cleanup of circularization product. To make DNA nanoball (DNB), the library is incubated at 30 C for 10 min using DNB enzyme. Finally, Library was quantified by QauntiFluor ssDNA System (Promega). Sequencing of the prepared DNB was conducted on the MGIseq system (MGI) with 150 bp paired-end reads.