Journal: Journal of visualized experiments : JoVE

Article Title: A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe

doi: 10.3791/59133

Figure Lengend Snippet: Materials

Article Snippet: RNase A/T1 mix Thermo Fisher Scientific EN0551 Use according to manufacturer recommendation Sterile Polystyrene Inoculating Loop Corning, Inc. OS101 Or equivalent, use to transfer colonies from agar plates to 96- well plate Sterile workspace and burners Tissue Culture Plate, 96-well Optical Flat Bottom with Low Evaporation Lid Corning, Falcon C353072 Or equivalent, must have optical flat bottom for micro-plate ready TruSeq DNA PCR-Free LT/HT Library Prep Kit Illumina, Inc. 20015962 Use to prepare the whole-genome sequencing library Yeast Extract, Powder Fisher Chemical BP1422–500 Use at 5 g/L to make liquid and solid rich media (YEA) Open in a separate window Materials For shearing parameters during the library preparation, use a focused sonicator (see ) and set the duty factor to 20%, peak power to 175 W, with 200 cycles per burst, and frequency sweeping mode at 5.5oC to 6 °C for 45 s. Alternatively, use a DNA and chomatin shearing system (see ) with the following settings: 50% amplitude at 4 °C with pulse mode, 15 s on and 15 s off for 10 min, with a total processing time of 20 min.

Techniques: DNA Extraction, Software, Evaporation, Sequencing