ID   CA554628   standard; RNA; EST; 490 BP.
XX
AC   CA554628;
XX
SV   CA554628.1
XX
DT   20-NOV-2002 (Rel. 73, Created)
DT   20-NOV-2002 (Rel. 73, Last updated, Version 1)
XX
DE   C0888B04-5N NIA Mouse Blastocyst cDNA Library (Long) Mus musculus cDNA
DE   clone NIA:C0888B04 IMAGE:30033423 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   1-490
RA   Piao Y., Kargul G.J., Dudekula D.B., Qian Y., Tanaka T., Luo A., Ko M.S.H.;
RT   "Systematic Analyses of NIA Mouse Blastocyst cDNA Library (Long)";
RL   Unpublished.
XX
CC   Other_ESTs: C0888B04-3
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: C0888  row: B  column: 04
CC   Seq primer: M13 Reverse
CC   High quality sequence stop: 490
CC   POLYA=No.
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..490
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 20 Blastocysts. Double-stranded
FT                   cDNAs were synthesized with an Oligo(dT) primer
FT                   [Invitrogen:
FT                   5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from
FT                   0.2 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:C0888B04 IMAGE:30033423"
FT                   /clone_lib="NIA Mouse Blastocyst cDNA Library (Long)"
FT                   /tissue_type="Blastocyst"
FT                   /dev_stage="3.5-dpc"
FT                   /lab_host="DH10B"
XX
SQ   Sequence 490 BP; 115 A; 121 C; 134 G; 120 T; 0 other;
     cgtggtcggg atcctgtgaa gcgggaagcc actctacctc cagtgagccc tccaaaggct        60
     gcgctgtctg tggatgaggt ggagaagaag tctaaggcca tcattgagga atatctccat       120
     ctcaatgaca tgaaggaggc agtacagtgt gtccaggagc tggcttcacc ctccttgctc       180
     ttcatcttcg tacggcttgg tatcgagtcc acgctggagc gtagtaccat tgctcgtgag       240
     catatggggc gactactgca ccaactgctc tgtgcggggc acctctctac tgcccagtac       300
     tatcaagggc tgtatgaaac actagaattg gctgaggaca tggaaattga tatccctcat       360
     gtatggcttt acctggcaga actgattaca cctattctcc aggaagatgg ggtgcccatg       420
     ggagagctct tcagggaaat tacgaagcct ctgagaccca tgggcaaagc tacttcttta       480
     ttgctggaga                                                              490
//