ID   CA545128   standard; RNA; EST; 106 BP.
XX
AC   CA545128;
XX
SV   CA545128.1
XX
DT   20-NOV-2002 (Rel. 73, Created)
DT   20-NOV-2002 (Rel. 73, Last updated, Version 1)
XX
DE   K0108E11-5N NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit+/Sca-1-) cDNA
DE   Library (Long) Mus musculus cDNA clone NIA:K0108E11 IMAGE:30040186 5', mRNA
DE   sequence.
XX
KW   EST.
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OS   Mus musculus (house mouse)
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Rodentia; Sciurognathi; Muridae; Murinae; Mus.
XX
RN   [1]
RP   1-106
RA   Piao Y., Kargul G.J., Dudekula D.B., Qian Y., Luo A., Carter M.G., Taub D.,
RA   Longo D.L., Keller J., Ko M.S.H.;
RT   "Systematic Analyses of NIA Mouse Hematopoietic Stem Cell
RT   (Lin-/c-Kit+/Sca-1-) cDNA Library (Long)";
RL   Unpublished.
XX
CC   Other_ESTs: K0108E11-3
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: K0108  row: E  column: 11
CC   Seq primer: M13 Reverse
CC   High quality sequence stop: 106
CC   POLYA=No.
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..106
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                   obtained from Drs. Dennis Taub, Dan Longo (National
FT                   Institute on Aging, USA), Jonathan Keller (National Cancer
FT                   Institute, USA). Double-stranded cDNAs were synthesized
FT                   with an Oligo(dT) primer [Invitrogen:
FT                   5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from
FT                   2.4 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/6NCr"
FT                   /clone="NIA:K0108E11 IMAGE:30040186"
FT                   /clone_lib="NIA Mouse Hematopoietic Stem Cell
FT                   (Lin-/c-Kit+/Sca-1-) cDNA Library (Long)"
FT                   /tissue_type="Hematopoietic Stem Cell (Lin-/c-Kit+/Sca-1-
FT                   )"
FT                   /dev_stage="Age approx.10 weeks old"
FT                   /lab_host="DH10B"
XX
SQ   Sequence 106 BP; 28 A; 32 C; 22 G; 24 T; 0 other;
     caataccaca tctgttatca agattgccgt ggagccggtt aacccctcag agctgcccaa        60
     aatgcttgat ggcctgcgca aggtcaacaa gagctatcca tctctc                      106
//