GSM1211029: WT_SOLID_310; Schizosaccharomyces pombe; OTHER
Based on Milman et al. 2009 Mol Cell Biol 29: 5998-6005 and Pan et al. 2011 Cell 144:719-731. Whole cell extracts were prepared by grinding cells with glass beads and extracting in 10%TCA. Pellets were then extracted with SDS and soluble protein was subjected to immunoprecipitation of Rec12-flag with anti-flag antibodies. Protein was digested with proteinase K and Rec12-associated oligonucleotides were recovered by ethanol precipitation. Based on Pan et al. 2011 Cell 144:719-731. Oligonucleotides were tailed with ribo-G using rGTP and terminal transferase, then ligated to duplex DNA adaptors with a dC overhang using T4 RNA ligase 2. Complementary strands were synthesized with Klenow, then purified by denaturing polyacrylamide electrophoresis. Purified complementary strands were subjected to 3' tailing with ribo-G using rGTP and terminal transferase, then another duplex adaptor was ligated using T4 RNA ligase 2. Ligated material was amplified with 16 cycles of PCR to add 454 or SOLiD adapters.
GEO
GSM1211029