characteristics.text: LIBEST_008727
characteristics.characteristic: External Id
characteristics.text: SAMN00163252
characteristics.characteristic: External Id
characteristics.text: 2014-02-03T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2020-06-04T21:03:09.307Z
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: Construction of this library was carried out using Stratagene's Lambda ZAP II-cDNA synthesis kit. cDNA was synthesized using an oligo dT primer and made blunt-ended. EcoRI adapters then were ligated to the blunt ends, excess adapters were removed, and the cDNA was ligated to Lambda ZAP II arms. The vector is designed containing the pBluescript sequence as well as lambda DNA and cDNA is cloned within this pBluescript sequence. The vector was then packaged using Gold Gigapackaging extracts. Library was grown in XLIBlue MRF' cells and amplified. The library was excised by mass excision using Stratagene's 'Mass excision kit' that uses exassist as a helper phage that releases the pBluescript sequence and circularises it as single strand plasmids that are then packaged (by helper phage) and secreted out of the host cell as phagemids. SOLR cells were transformed with phagemids and the library was plated out on LB-amp plates to select for transformants. Approximately 500,000 colonies were plated and pooled for isolation of the double stranded plasmid DNA library. This DNA was used to transform DH10B cells, the clones from which were used as a source of DNA for the sequencing project. This library was constructed by Dr. Stavros Bashiardes as part of the Physcomitrella EST program (PEP) at the University of Leeds (UK) and Washington University in St. Louis (USA). DNA sequencing by Washington University Genome Sequencing Center. For information on obtaining a clone please contact: Celia Knight (c.d.knight@leeds.ac.uk).
characteristics.characteristic: description
characteristics.text: DH10B
characteristics.characteristic: lab-host
characteristics.text: Physcomitrium patens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3218
characteristics.characteristic: organism
characteristics.text: EcoRI
characteristics.characteristic: re_1
characteristics.text: EcoRI
characteristics.characteristic: re_2
characteristics.text: protonemata 7 day old, abscisic acid treated
characteristics.characteristic: tissue-type
characteristics.text: Moss EST library PPA
characteristics.characteristic: title
characteristics.text: phagemid
characteristics.characteristic: v_type
characteristics.text: pBluescript SK-
characteristics.characteristic: vector