Amplicon-seq of nad9 locus from Nicotiana tabacum, point mutated at nad9, amino exchange Q89K - Experiment 2
After initial quality control using Agilent's Bioanalyzer sequencing libraries were prepared from 50ng of DNA per sample following Roche's  "KAPA DNA Hyper Prep Kit" library preparation protocol for double indexed Illumina libraries: First, end repair and A-tailing were performed. Next, Illumina sequencing compatible adapters carrying unique dual indices were ligated (NEXTFLEX Unique Dual Index Barcodes).  Following bead-based clean-up steps the libraries were amplified using 5 cycles of PCR. Library quality and size was checked with qBit, Agilent Bioanalyzer and qPCR. Sequencing was carried out on an Illumina MiSeq Micro flow cell in PE150bp mode yielding between 140,000-200,000 fragments per sample.