Journal: BMC Genomics
Article Title: Genotyping 1000 yeast strains by next-generation sequencing
Figure Lengend Snippet: Library preparation pipeline. DNA isolation is performed with a 96-head liquid handling robot. DNA fragmentation is achieved by sonication, either in glass tubes (Covaris) or PCR strips (Bandelin). SPRI bead cleanup is automated on a 96-head liquid handling robot. Three enzymatic steps for barcoded adapter ligation are performed by addition of enzyme (+ buffer), incubation, and heat inactivation in a thermocycler. After pooling of 48 barcoded libraries, samples are concentrated and size-selected using an E-gel. PCR is performed on the size-selected pool to enrich for adapter containing fragments and elongate them to full-length libraries. A final cleanup is performed in PCR strips mounted to a homemade magnetic stand.
Article Snippet: DNA purification with SPRI beads The amplified DNA was purified in 0.2 ml PCR strips by mixing the DNA with 1x volume of Agencourt AMPure XP beads (Beckman Coulter) and select the magnetic beads with a homemade magnetic stand (Additional file : Figure S6).
Techniques: DNA Extraction, Sonication, Polymerase Chain Reaction, Ligation, Incubation