characteristics.text: LIBEST_023632
characteristics.characteristic: External Id
characteristics.text: SAMN00166525
characteristics.characteristic: External Id
characteristics.text: Centre for Environmental Stress and Adaptation Research, Bio21 Institute, Genetics Department, University of Melbourne
characteristics.characteristic: INSDC center name
characteristics.text: 2008-08-29T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:46:20.837Z
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 using Trizol (Invitrogen) and messenger RNA was isolated using Nucleotrap mRNA kit (Macherey-Nagel). First strand cDNA was prepared using the Creator SMART cDNA Library Construction Kit (Clontech) with two modifications: a modified 3' primer (5'-AAGCAGTGGTATCAACGCAGAGTGGCCGAGGCGGCCTGTTTTGTTTTTTTTTCTTTTTTTTTTVN-3') and a 1:1 mixture of ArrayScript Reverse Transcriptase (Ambion) and BioScript Reverser transcriptase (Bioline). Second strand cDNA was synthesised using the Advantage 2 PCR System (Clontech) with adapter-specific primer (5'-AAGCAGTGGTATCAACGCAGAGT-3'). Double stranded cDNA was purified using the DNA Clean And Concentrator (Zymogen) and digested with Sfi-I restriction enzyme (New England Biolabs). Digested cDNA was purified and ligated to pDNR-Lib vector (Clontec) and heat transformed into JM109 (Promega). 576 random colonies were picked and the cDNA inserts were amplified using M13 primers. Unincorporated nucleotides and oligos were removed using ExoZap (Stratagene) and the amplicons were sequenced from the M13F end.
characteristics.characteristic: description
characteristics.text: adult
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001272
characteristics.characteristic: dev-stage
characteristics.text: JM109
characteristics.characteristic: lab-host
characteristics.text: Micropterix calthella
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_41027
characteristics.characteristic: organism
characteristics.text: Sfi-I A
characteristics.characteristic: re_1
characteristics.text: Sfi-I B
characteristics.characteristic: re_2
characteristics.text: female and male
characteristics.characteristic: sex
characteristics.text: whole body
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/BTO_0001489
characteristics.characteristic: tissue-type
characteristics.text: Micropterix calthella adult whole body cDNA library
characteristics.characteristic: title
characteristics.text: plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pDNR-Lib
characteristics.characteristic: vector