QIAprep 96 Turbo Miniprep Kit (24)

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Publication protocol

Glaucocystis nostochinearum total RNA was extracted after breaking of cells with glass beads [S1], with TRIZOL (Invitrogen) instead of the extraction buffer and following the manufacturer’s instructions for the RNA purification steps. A cDNA library was con- structed with 100 ng polyA􏰀 RNA (isolated with Oligotex-dT Cellu- lose Type 7; Amersham Biosciences; Piscataway, NJ) and primers, plasmid pDNRlib, and reverse transcriptase from the CreatorTM SmartTM cDNA Library Construction kit (BD Biosciences Clontech; Palo Alto, CA). Plasmids were isolated from bacterial cultures with the QIAprep 96 Turbo Miniprep Kit (Qiagen), and sequencing reac- tions were performed with the ABI Prism BigDyeTM Terminators version 3.0/3.1 (Perkin-Elmer, Wellesley, MA). The purified sequenc- ing reactions were separated and analyzed either on an ABI 3700 (Cyanophora ESTs) or an MJ BaseStation automatic sequencer (Glaucocystis ESTs).

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