characteristics.text: 2011-11-09T17:00:40Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-09T09:53:18Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA1026959
characteristics.characteristic: External Id
characteristics.text: ED
characteristics.characteristic: INSDC center name
characteristics.text: 2011-11-09T17:00:40Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-09T09:53:18Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS023775
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-540:Synthetic 10000 DNA library
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: PCR read-off microarrays: Extension reaction mix (200 uL) without primers was prepared according to a Promega standard protocol using a PCR Master Mix (Promega; 25 U/mL Taq Polymerase, 200 uM dNTP, 1.5 mM MgCl2) was loaded onto the microarray using an Agilent hybridization cover slide. The first extension step was carried out at 55C for 16 h (overnight). Hereafter, the reaction mixture was removed using a pipette and fresh PCR reaction mix (200 uL) with Solexa-primer-1 and 2 (0.1 uM) were loaded onto the microarray and a standard PCR cycle was set up in a GeneMachines Hyb4 automated hybridizer [40 cycles; denaturation at 94C for 30 s, annealing at 65C for 1 min, extension at 70C for 1 min]. In addition, an initial 3 min denaturation step at 94C and a final 15 min extension step at 70C were carried out. Immediately after the PCR had finished the reaction mix was collected using a pipette and the microarray washed with H2O (3 x 50 uL). The aqueous fractions were pooled together and concentrated by speed vac followed by purification by preparative DNA gel electrophoresis (dsDNA-10,000: 1.65 ug, 40% isolated yield). PCR in solution: The purified product (250 ng) from the PCR read-off microarrays were used as templates in another round of PCR with Solexa-primer-1 and 2 (1 uM) in a 1 x PCR Master Mix (200 uL) in a Techne TC-312 PCR cycler with the same cycle as on microarray. After PCR the samples were concentrated by speed vac followed by purification by preparative DNA gel electrophoresis (dsDNA-10,000-2: 16.8 ug, 30% isolated yield). This sample is not extracted from any biosample material as it is a synthetic PNA library.
characteristics.characteristic: description
characteristics.text: synthetic construct
characteristics.characteristic: organism
characteristics.text: synthetic DNA
characteristics.characteristic: organism
characteristics.text: synthetic construct
characteristics.characteristic: scientific_name
characteristics.text: Synthetic 10000 DNA library
characteristics.characteristic: title