LOCUS JK034125 753 bp mRNA linear EST 07-FEB-2014 DEFINITION 43 (LHA-2011) Bellamya aeruginosa normalized cDNA library Bellamya aeruginosa cDNA 5', mRNA sequence. ACCESSION JK034125 VERSION JK034125.1 DBLINK BioSample: SAMN00622379 KEYWORDS EST. SOURCE Sinotaia aeruginosa ORGANISM Sinotaia aeruginosa Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Mollusca; Gastropoda; Caenogastropoda; Architaenioglossa; Viviparoidea; Viviparidae; Sinotaia. REFERENCE 1 (bases 1 to 753) AUTHORS An,L.H. and Zheng,B.H. TITLE Construction and Characterization of Normalized cDNA Library from the Snail Bellamya aeruginosa Exposure to Copper JOURNAL Unpublished COMMENT Contact: An L.H. State Environmental Protection Key Laboratory Of Estuarine And Coastal Research Chinese Research Academy For Environment Sciences 8 Dayangfang BeiYuan Road, Chaoyang District, Beijing 100012, China Tel: +86-10-84913914 Fax: +86-10-84913914 Email: anlh@craes.org.cn PCR PRimers FORWARD: CTCGGGAAGCGCGCCATTGTGTTGGT BACKWARD: ATACGTCTCACTATAGGGCGAATTGGCC. FEATURES Location/Qualifiers source 1..753 /organism="Sinotaia aeruginosa" /mol_type="mRNA" /db_xref="taxon:2653872" /tissue_type="gonads; digestive gland; peripheral tissue" /clone_lib="SAMN00622379 Bellamya aeruginosa normalized cDNA library" /dev_stage="mature" /note="Vector: Lambda TripEx2; 1.5 microliter RNA solution (1-3 microgram of total RNA) was normalized using CDS and SMART Oligo IV primer firstly, and then the reaction solution was reversed transcription for synthesizing the first-strand using MMLV reverse transcriptase (Promega, USA). Subsequently, PCR reaction was operated and cDNA was extracted using Promegas PCR clean-up system kit (Madison, USA). And then the cDNA library was constructed using SMART cDNA Library Construction Kit. After that, 6000 random clones were sequenced by the ABI3730xl. BLAST homology searches in the nr and nt databases in GenBank were performed for each of the unigenes under Evalues of 1e-5. The unigenes were named after the homologous sequences in GenBank. The annotated unigenes were then matched to specific processes or functions using COG and GO tools." BASE COUNT 208 a 125 c 135 g 285 t ORIGIN 1 ttcatttact gacaaaatct attgagtgtt ttgggaagaa ttggctgcca tgacttcaga 61 gtctttgatg atggggcttt tacttatctc ttgtgtcagt gctattttct cggcatcaac 121 tggggagcct gctcagtgct tcccgcaaaa tgcagagtgt gacccactca ttaactttgg 181 gacccctaac aattgctgtg cagacttcca aagatgtgtt gcccattctg gactggatgg 241 aaaagtaact tacaggtgta cctcaattta cattggatag actcaagagg cactgatctt 301 tgaagcagtg ttgttttata tcagtctatg atcgaatact gttttttgct tattttttgc 361 tttgattctt actattgatt tatgcctatc atttatttgc acataaagag gtttcttatt 421 attatgccta ccattttatt ttgcaatcaa gagatttctt agtattgatt tatacctgtc 481 atttagtgtg tatctaaagt agatctccca agattcaata taatctacag ctgaatatta 541 ttgacttaag ataaatctcc cagcattgat gtacacttgc attgattgta ctatttaatg 601 ctctgtgtcc taagtgatca gccaagattc aatgttaaat gcttcaaagg taatataaat 661 aaatcataaa atgtatttgg ggttaattca ttgggttctt aacttagtct tttgaaagtc 721 agattttatg aaaaagtttc ttttatatat gac //