characteristics.text: SAMN00166461
characteristics.characteristic: External Id
characteristics.text: LIBEST_023568
characteristics.characteristic: External Id
characteristics.text: Chang Gung University
characteristics.characteristic: INSDC center name
characteristics.text: 2009-12-31T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:46:18.887Z
characteristics.characteristic: INSDC last update
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Generic
characteristics.characteristic: NCBI submission model
characteristics.text: Generic.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: Total RNA was extracted from mid-logarithmic phase P. hominis trophozoites by using TRI REAGENT (Molecular Research Center, Inc.) following the manufacturers' protocols. The integrity and quality of the extracted total RNA was assessed by agarose gel electrophoresis and spectrophormetry. PolyA+ mRNA was isolated from 1 mg of total RNA using polyATtract mRNA isolation system IV (Promega, Madison, USA). Approximately 0.5 ugg of polyA+ mRNA was used for the construction of a directional cDNA library construction by using a cDNA Synthesis Kit (Stratagene, Cedar Creek, USA) following the manufacturer's protocol. The cDNA was size fractionated using by agarose gel electrophoresis followed by purification using Qiaex II Gel extraction kit (Qiagen). Purified cDNA was ligated into EcoRI/XhoI digested pBluescript II XR (Stratagene) using temperature-cycle ligation followed by PCR validation of the ligation reaction. The ligation product was precipitated and electro-transformated into E. coli XL1-Blue MRF' (Stratagene) and plated on LB agar containing IPTG and X-Gal for blue/white selection. Positive clones containing an insert were picked into 2xTY (100 ugg/ml Amp, 10% [vol/vol] glycerol) in 96-well plates and incubated for 24 hours at -37oC for plasmid purification. The insert length of each library was evaluated in 96 clones by PCR. T3 primer was used from the polylinker of the vectors to sequence the 5' end of each insert using standard protocols.
characteristics.characteristic: description
characteristics.text: Mid-log phase trophozoite
characteristics.characteristic: dev-stage
characteristics.text: Pentatrichomonas hominis
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_5728
characteristics.characteristic: organism
characteristics.text: XhoI
characteristics.characteristic: re_1
characteristics.text: EcoRI
characteristics.characteristic: re_2
characteristics.text: ATCC30000
characteristics.characteristic: strain
characteristics.text: Ph30000 cDNA Library
characteristics.characteristic: title
characteristics.text: Plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: XL1-Blue MRF'
characteristics.characteristic: vector