Project name: Lachancea kluyveri strain:CBS 3082

Description: Lachancea kluyveri CBS 3082 (NRRL Y-12651 diploid type strain) was used as the ancestral strain to generate the evolved strains. A single colony from L. kluyveri (CBS 3082) was evolved by a long-term modified serial dilution transfer method (batch cultures) method in which bacteria were sequentially introduced to compete with yeasts under aerobic conditions. Genomic DNA of ancestor and evolved strains of L. kluyveri was extracted using methods previously described (Philippsen et al. 1991). Concentrations of purified nucleotides were determined at 260 nm using a NanoDrop 2000 spectrophotometer (Thermo Scientific) and purity assessed at an absorbance ratio of 260/280 nm and 260/230 nm. DNA fragment size was determined using Bioanalyzer 2100 (Agilent Technologies). Genomic DNA was then fragmented to an average size of 550 bp using a Covaris M220 (Covaris). Fragments were subsequently end repaired, A-tailed, indexed adapters and ligated. The products were purified and enriched with PCR to create the final cDNA library. The 2 tagged cDNA libraries were pooled in equal ratios and used for 2 × 100 bp paired-end sequencing on a single lane of the Illumina MiSeq (Illumina) using MiSeq Reagent kit v2. at the core sequencing facility of Academia Sinica, Taiwan.

Data type: genome sequencing

Sample scope: Monoisolate

Relevance: Evolution

Last updated: 2015-01-20