ID   CA539541   standard; RNA; EST; 493 BP.
XX
AC   CA539541;
XX
SV   CA539541.1
XX
DT   20-NOV-2002 (Rel. 73, Created)
DT   20-NOV-2002 (Rel. 73, Last updated, Version 1)
XX
DE   C0280B10-5N NIA Mouse 7.5-dpc Whole Embryo cDNA Library (Long) Mus musculus
DE   cDNA clone NIA:C0280B10 IMAGE:30018261 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   1-493
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RL   Unpublished.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   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: C0280  row: B  column: 10
CC   Seq primer: M13 Reverse
CC   High quality sequence stop: 493
CC   POLYA=No.
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..493
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:C0280B10 IMAGE:30018261"
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"
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SQ   Sequence 493 BP; 133 A; 101 C; 81 G; 178 T; 0 other;
     ccctaacgta agaaatacgc tgcaaaatat cgagtaataa agaggccacc atattttaac        60
     acccctaccc ccccccattt catatcactt ttctgtgtgc gtcgctctga acttttctct       120
     ctaaagacta aaaaaatacc tcttaacgtg aacgcttctc caaagaagat accaagtcca       180
     tatgtatcac gtaattattt tccttttatg tctggttctc tacatttttc taaaggtaga       240
     tattactagt attttcatct taattttagg ctgaagatga acccgagcct gagccccatc       300
     ctgaaaatgt tagctgttgc aggtggagcg tctttcatat gtccctgggc tgcagcattt       360
     tgttctacta aagttttttt gtttttttgt ttttcctatt gattatgagg atggggaaga       420
     actgactgca ggtagattaa catcatttac aaaaggatat agagtttttc aatttctcat       480
     gttttttctt ttg                                                          493
//