characteristics.text: LIBEST_022675
characteristics.characteristic: External Id
characteristics.text: SAMN00165191
characteristics.characteristic: External Id
characteristics.text: American Museum of Natural History
characteristics.characteristic: INSDC center name
characteristics.text: 2009-02-15T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:51.723Z
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: Around 200 male Drosophila mayaguana flies (stock 15081-1397.03 from the Tucson Stock Center) were dissected for their accessory glands. Dissections were done in PBS with RNA later (Ambion). Accessory glands were pooled together and kept in RNA later at -20oC until RNA extraction. Total RNA was extracted with the RNAeasy extraction kit (QIAGEN) and mRNA was isolated from total RNA using the Oligotex mRNA kit (QIAGEN). cDNA synthesis was done with the Super SMART cDNA Synthesis Kit (Clontech). Female cDNA generated by the same process was used as probe for subtraction of housekeeping genes and other non-male specific genes using the PCR-Select cDNA Subtraction Kit (Clontech). The last PCR product of the subtraction process was size fractionated for fragments larger than ~700bp. Size fractionation was carried out in 2% low-melting agarose gel stained with SYBR. PCR fragments were retrieved from the gel using the Wizard gel purification kit (PROMEGA). After a quick incubation with Taq polymerase for adding overhanging A's, the retrieved fragments were cloned into a TA plasmid vector (Topo TA cloning kit, Invitrogen). Chemically competent cells (Invitrogen) were then transformed to produce an accessory gland subtracted library.
characteristics.characteristic: description
characteristics.text: adults
characteristics.characteristic: dev-stage
characteristics.text: E. coli
characteristics.characteristic: lab-host
characteristics.text: accessory gland
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0010147
characteristics.characteristic: organ
characteristics.text: Drosophila mayaguana
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_7268
characteristics.characteristic: organism
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: Tucson Stock Center 15081-1397.03
characteristics.characteristic: strain
characteristics.text: Accessory gland cDNA library
characteristics.characteristic: title
characteristics.text: plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pCR2.1-TOPO
characteristics.characteristic: vector