ID   CA540942   standard; RNA; EST; 511 BP.
XX
AC   CA540942;
XX
SV   CA540942.1
XX
DT   20-NOV-2002 (Rel. 73, Created)
DT   20-NOV-2002 (Rel. 73, Last updated, Version 1)
XX
DE   C0298E07-5N NIA Mouse 7.5-dpc Whole Embryo cDNA Library (Long) Mus musculus
DE   cDNA clone NIA:C0298E07 IMAGE:30020022 5', mRNA sequence.
XX
KW   EST.
XX
OS   Mus musculus (house mouse)
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Rodentia; Sciurognathi; Muridae; Murinae; Mus.
XX
RN   [1]
RP   1-511
RA   Piao Y., Kargul G.J., Dudekula D.B., Qian Y., Tanaka T., Lim M.K., Luo A.,
RA   Ko M.S.H.;
RT   "Systematic Analyses of NIA Mouse 7.5-dpc Whole Embryo cDNA Library
RT   (Long)";
RL   Unpublished.
XX
CC   Other_ESTs: C0298E07-3
CC   Contact: Dawood B. Dudekula
CC   Laboratory of Genetics
CC   National Institute on Aging/National Institutes of Health
CC   333 Cassell Drive, Suite 4000, Baltimore, MD 21224-6820, USA
CC   Email: cdna@lgsun.grc.nia.nih.gov
CC   Plate: C0298  row: E  column: 07
CC   Seq primer: M13 Reverse
CC   High quality sequence stop: 511
CC   POLYA=No.
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..511
FT                   /db_xref="taxon:10090"
FT                   /note="Vector: pSPORT1 (Invitrogen); Site_1: SalI; Site_2:
FT                   NotI; Mouse cDNA project by the Laboratory of Genetics,
FT                   National Institute on Aging (NIA), Intramural Research
FT                   Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a
FT                   long-transcript enriched cDNA library (Ref. Genome Res. 11:
FT                   1553-1558 (2001). [PMID: 11544199]). Total RNAs were
FT                   extracted from a pool of four embryos at 7.5-days
FT                   postcoitum. Double-stranded cDNAs were synthesized with an
FT                   Oligo(dT) primer [Invitrogen:
FT                   5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 7
FT                   ug of total RNA, treated with T4 DNA polymerase, and
FT                   purified by ethanol-precipitation. The cDNAs were ligated
FT                   to Lone-linker LL-Sal4, purified by phenol/chloroform, and
FT                   separated from free linkers by Centricon 100. Then, the
FT                   cDNAs were amplified by long-range high fidelity PCR using
FT                   Ex Taq polymerase (Takara) with a primer Sal4-S. The
FT                   products were purified by phenol/chloroform and Centricon
FT                   100. The cDNAs were digested with SalI and NotI enzymes and
FT                   cloned into SalI/NotI site of pSPORT1 plasmid vector. The
FT                   DH10B E. coli host was transformed with the ligation
FT                   mixture by the standard chemical method. The average insert
FT                   size is about 2.2 kb. The library was constructed by Yulan
FT                   Piao (NIA)."
FT                   /organism="Mus musculus"
FT                   /strain="C57BL/6J"
FT                   /clone="NIA:C0298E07 IMAGE:30020022"
FT                   /clone_lib="NIA Mouse 7.5-dpc Whole Embryo cDNA Library
FT                   (Long)"
FT                   /tissue_type="whole embryo including extraembryonic tissues
FT                   at 7.5-days postcoitum"
FT                   /dev_stage="7.5-days postcoitum"
FT                   /lab_host="DH10B"
XX
SQ   Sequence 511 BP; 145 A; 93 C; 142 G; 131 T; 0 other;
     ggcagtgtgg acctctgtga tgggacatct tgtgggatct cacagccagt tctgtggcaa        60
     aggaggatga aggcttacga ggtgagtact gaatagtaat gagaagttag gttgaaagga       120
     cagtgccaca atgatgacat catatttgct actcttgaca gctggggtga cgatagcttt       180
     agaccaccat gattataact gaggcacaca aaataatgat ttgggggcta cttacagagt       240
     aagcagtttt tacataaaga ttcccccccc ccaggactcg tcaggaagct ggataacaga       300
     gcgagcgcaa tgtaagcaat tttgtgtgtg catgtggtcc cattacaagg aatacccaat       360
     ggcaaatgag cactaaagat ccctcaaccc acacattgaa tgtattgttc cctgagagca       420
     attgtagata gaggagtggg tgggaagtct gaagtcaggg cattgtttct gcgtatctga       480
     ggaggctggt ttttgatatg tgggagctag t                                      511
//