title: green berries 2009 for 454 sequencing
organism: Vaccinium corymbosum
description: Two libraries were prepared from samples of green and ripe fruit respectively from plants of the O'Neal variety of southern highbush blueberry Vaccinium corymbosum. The green fruit library was prepared from a mix of unripe, green fruits of varying sizes harvested on May 18, 2009 from plant 2-41. The ripe fruit library was prepared from ripe fruits harvested on June 15, 2009 from plants 2-40, 2-41, and 2-42, also of the O'Neal variety. The libraries for sequencing were constructed using the SMART PCR cDNA synthesis kit from CloneTech. The 3' and 5' primers used in first strand cDNA synthesis were aagcagtggtatcaacgcagagtact(30)VN  and aagcagtggtatcaacgcagagtacgcggg, respectively, where V was a,g, or c and N was  any nucleotide.  The products of first strand cDNA synthesis were amplified using a PCR polyA disruption primer designed to introduce non-A bases in the polyA tail region of the cDNA, since homopolymeric sequences are difficult to sequence using the 454 technology. The polyA disruption primer sequence was attctagaggccgaggcggccgacatgt(4)gtct(4)gttctgt(3)ct(4)VN, where numbers in parenthesis indicate the number of times the preceding base appeared in the sequence and V was a,g, or c. The sequence of the five prime primer used for the second strand cDNA synthesis was aagcagtggtatcaacgcagagt. The two libraries were sequenced in two sectors of the same plate on a 454-GS FLX Titanium sequencer (454 Life Sciences, Roche Diagnostics, USA) at the David H. Murdock Research Institute. 
attributes.tag: cultivar
attributes.value: O'Neal
attributes.tag: biomaterial_provider
attributes.value: Ketan Patel
attributes.tag: tissue
attributes.value: berry
attributes.tag: collected_by
attributes.value: Ketan Patel
attributes.tag: collection_date
attributes.value: 5/18/09
attributes.tag: dev_stage
attributes.value: green
attributes.tag: growth_protocol
attributes.value: field
attributes.tag: N
attributes.value: 1
attributes.tag: geo_loc_name
attributes.value: missing
attributes.tag: BioSampleModel
attributes.value: Plant