characteristics.text: SAMN00163251
characteristics.characteristic: External Id
characteristics.text: LIBEST_008726
characteristics.characteristic: External Id
characteristics.text: Washington University School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2001-03-08T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2020-06-04T21:03:09.279Z
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 performed by Dr. W. Gregg Clark using a modification of the cDNA synthesis protocol developed in the laboratory of Dr. Michael Lovett by Dr. Yulia Korshunova (personal communication). First polyA + RNA was isolated from total gametophore RNA using oligo dT magnetic beads. Following this, first strand cDNA synthesis was performed on the bead-bound polyA + RNA, during which an oligonucleotide anchor sequence was incorporated onto the 5'-ends of the cDNA. PCR amplification was then used to synthesize the second strand, to amplify the double stranded DNA, and to incorporate dUTP containing sequences into the ends of the double stranded cDNA. This DNA was size selected and cloned into pAMP1 using the CloneAMP pAMP1 System (Life Technologies, GibcoBRL) for cloning amplification products by a non-restriction site dependant process. The cloning was directional based on sequence asymmetry introduced at the ends during PCR amplification. The 3' cDNA ends are proximal to the NotI site of the multiple cloning site in pAMP1. This annealing mixture was transformed into chemically competent DH10B cells and selected for ampicillin resistant growth. The resulting clones (about 330,000) were pooled to make the library.
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: gametophore: 30 day old tissue, ammonium-grown
characteristics.characteristic: tissue-type
characteristics.text: Moss EST library PPG
characteristics.characteristic: title
characteristics.text: pAMP1
characteristics.characteristic: vector