GrainGenes Sequence Report: Ta.8319.1.A1

Locus Comment: Contact: Wm L Crosby; Bioinformatics; University of Saskatchewan, Department of Computer Science; 1C101 Engineering Building, 57 Campus Drive, Saskatoon,; Saskatchewan, S7N 5A9, Canada; Tel: 306 966 1769; Fax: 306 966 2033; Email: fgas_ests@cs.usask.ca; This sequence is the direct result of the Base calling software; Phred (default parameters). It is the raw base calls. To aid in the; identification of the high quality insert the software Lucy; (default parameters) has been run on this sequence. Lucy identified; the region [12,619].; Plate: L6B018 row: L column: 20.

Note: Organ: Crown and leaf; Vector: pCMV.SPORT6; Crown (50%) and leaf (50%) tissues from wheat cultivar Norstar after short exposure times to low temperature in the light and in the dark. 12 mRNA populations were combined before constructing the library. The first 6 populations: After 7 days of growth at 20Cs from wheat cultivar Norstar after short exposure times to low temperature in the light and in the dark. 12 mRNA populations were combined before constructing the library. The first 6 populations: After 7 days of growth at 20, wheat plants were transferred to 4C in the light. 1cm crown sections and green leaf tissu were separately harvested after 1, 3, and 6 hours of low temperature exposure. The last 6 populations: After 7 days of growth at 20C, wheat plants were transferred to 4C in the dark. 1cm crown sections and green leaf tissu were separately harvested after 1, 3, and 6 hours of low temperature exposure. First strand synthesis in this library was done in the presence of methylated dCTP thereby protecting from internal cleavage with NotI. In addition, this library used a primer for second strand synthesis that annealed to an artificial sequence (RNA oligo) added before first strand synthesis. Therefore when sequences from EST generated from this library will be masked for vector and adaptor sequences, an additional masking step will have to be included to mask this RNA oligo that is common to all clones (sequence CGACTGGAGCACGAGGACACTGACATGGACTGAAGGAGTAGAAA).

DNA

agagcggccgccctttttttttttttttttttttttttttgggatcaacc
gaccgtccatgaataaaacagtacacgtgggcacagtacaaatgaaatct
tcagcggaacaaccgaggcaaacccgggcgtaagctaataagagaaccta
caggattgtatgttgtatgcactattgttctgaaaatctgcaaactctat
tacattaggccgggtcgagaatggaaggggaggcggggcgacgccatggc
ggcccggtcaaggttgctccttttcttctctttcccttccttctttacgg
cttgaggctgcttcttcttggcgccatcgctggaatctttagcggacggc
gagggcgaggaagccaaggagttgttcttggggagcaccttgacatccga
acacccacagcaaaaccccattgatgctctgctaagcttctacaccacta
aaatttccccgtccctctctttctttctttctttcctctgctctccaagc
ttagagaaaactcaggtcttctctgctactgatcgtgtgtcagtgtgtgc
ttatgtgtttctgttttctactccttcagtccatgtcagtgtcctcgtgc
tccagtcgcggacgcgtgggtcgacgatatcccgggaattccggaccggt
accagcctgcttttttgtacaaacttgttctatagtgtcacctaaatagg
cctaatggtcatagctgttccctgtgtgaaattgttatccgtcccgcggc
ctaggctaaagtccggaggctgaatcggtcccgggtgtcttctatggagg
tcaaaaacaggtggatgggttctccaggcgatctgacgggtcacttaaac
aggctctgtttatattaaacttccaacgtaaacccctaccggccatttgg
gttcaatggggggcgggtttgtacgaaatttttgggaagtcccgctgaat
tttggggcccaaacaaaacccccatttgaccttaatggggtggaaaattt
tggaatcccccgtgaattaaaccgggtttcacgccccattggtttttctg
caaaaccggattcccgtggtaataggggaggcaaatacactaaagttccg
ccccactttgacaatccccataggg


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