title: 2bRAD-Seq of Chlamydomonas wild-collected line from repositories: Sample Cre_line_1690
design: "Chlamydomonas reinhardtii wild-collected isolates were obtained from the UTEX algae collection and from the Chlamydomonas Center, then maintained at University of Texas at Austin on tris-acetate-phosphate medium. gDNA was extracted from pools of each isolate using Qiagen DNeasy kits per manufacturer's protocol except no vortexing was used and the final elution was in 40ul NFW. 2b-RAD (Wang et al. 2012) libraries were prepared by first digesting each sample with AlfI and-or BcgI restriction enzymes (Fisher Scientific); next, SOLiD adapters were ligated with T4 ligase (NEB); and last, barcodes were incorporated with a 10-12 cycle PCR (98C x 5 sec., 60C x 20 sec., 72C x 10 sec.). Final products were selected at the 136-bp band by gel excision. Libraries were sequenced at the University of Texas Genome Sequencing and Analysis core facility on an ABI SOLiD 5500XL sequencer. Raw reads were filtered to remove low-quality (mean qual < 20) reads, then trimmed to 37 colorspace bases (for 36-bp reads, the length of the restriction enzyme cuts)."