metagenome_AnnaTempExp_2012-DNaseq-10degC-1
DNA was extracted from 50 mL of pelleted culture (Zhou et al., 1996). 2 μg of DNA were mechanically fragmented with a Nebulizer (Roche, 32 psi, 6 min in 500 μl nebulization buffer). Fragmented DNA was purified with a MinElute PCR purification column (Qiagen, Düsseldorf, Germany) and eluted in 50 μl low TE (Life Technologies, Darmstadt, Germany). The entire eluate was used to prepare a barcoded PGM sequencing library with the Ion Xpress plus gDNA Fragment Library Preparation Kit (manual Pub. No. 4471989, Rev M, May 2013) (Life Technologies). Library insert sizes were approximately 200 bp. The library was sequenced with a Personal Genome Machine (PGM) using the chemistry for 200 bp libraries.
2012-DNA-10-1