title: 454 sequencing of Coleochaete orbicularis transcriptome fragment library
design: 3-5ug of RNA was nebulized to a mean size range of 3-500bp. This is then followed by size selection of fragments >300bp by column exclusion and Ampuretm (Agencourt Bioscience)isolation.  The correct size selection was confirmed on an Agilent DNA 1000 LabChip.  Adapters were ligated onto the fragments and then fragments with correct adapters were selected using library capture beads.  The single stranded fragments were isolated with 0.125 N NaOH, followed by neutralization with acetic acid, and purified.  Single stranded library was validated qualitatively by Agilent RNA Pico 6000 LabChip and quantitatively by Invitrogen Ribogreen assay.  Standard library dilutions were made according to published protocol.