characteristics.text: LIBEST_002528
characteristics.characteristic: External Id
characteristics.text: SAMN00157087
characteristics.characteristic: External Id
characteristics.text: Washington University School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 1999-10-29T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2020-06-04T21:03:09.254Z
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 the cDNA library was carried out using Stratagenes 'UniZAP - cDNA synthesis kit'. cDNA was constructed using an oligo dT primer/linker that contains a XhoI site within it. Following ds cDNA synthesis, EcoRI adapters were ligated to the blunt ends and sample was digested with XhoI. The result is cDNA with an EcoRI sticky end on one side and a XhoI sticky end on the other. This cDNA was ligated directionally in UniZAP 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 Stratagens 'Mass excision kit' that uses exassist as a helper phage that releases the pBluescript sequence and circularises it as single stranded 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 1,000,000 colonies were grown and recovered. The double stranded plasmid library was recovered by using Quiagen Midi prep kit. 2 micro grams of each library were used to transform DH10B cells by electroporation.
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: XhoI
characteristics.characteristic: re_2
characteristics.text: protonemata: 7 day old tissue auxin treated
characteristics.characteristic: tissue-type
characteristics.text: Moss EST library PPN
characteristics.characteristic: title
characteristics.text: phagemid (ampicillin resistant)
characteristics.characteristic: v_type
characteristics.text: pBluescript SK-
characteristics.characteristic: vector