characteristics.text: SAMN00153250
characteristics.characteristic: External Id
characteristics.text: LIBEST_021544
characteristics.characteristic: External Id
characteristics.text: USDA-ARS
characteristics.characteristic: INSDC center name
characteristics.text: 2007-07-13T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:20.677Z
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: Experiments were carried out on sugarbeet root maggot, Tetanops myopaeformis Röder, field-collected diapausing and matured third-instar larvae, that had been maintained in laboratory storage at 6+1oC for one, two, and five years. Larvae were collected from fields near St.Thomas (SE quarter of section 25, 160o N and range 53 W), in Pembina County, North Dakota. Larvae were dug from the soil, placed into plastic bags containing loose field soil, and brought to the laboratory where they were washed and transferred into plastic storage bags filled with moist silica sand. Insects in bags were then stored in an incubator at 6+1oC. The larvae were in pre-diapause at the time of collection from the field. Therefore we can be reasonably confident that the insects used for the 1-, 2-, and 5-year assessments had entered diapause soon after collection. Total RNA was extracted from field-collected (November 2004) diapausing larvae and larvae that had been stored in the laboratory for one, two, and five years using Trizol reagent. Differential display was carried out according to the manufacturer's protocol using GeneFishingTM DEG Premix kits (Seegene, Rockville, MD, USA). Fragments were cloned into pCR2.1 and sequenced, and primers were designed to synthesize probes. For northern blot analysis the concentrations of RNA samples were determined using a spectrophotometer and ethidium bromide was added to all samples to ensure equivalent loading. Total RNA (5 ug/sample) was separated on a 1% agarose denaturing gel (0.41M formaldehyde, 1X MOPS-EDTA-sodium acetate). At the end of each gel run, the resulting gel was photographed to compare rRNA intensity. The RNA was subsequently transferred onto a positively charged nylon membrane with downward capillary action using 20X SSC (i.e., 3 M NaCl, 0.3 M sodium citrate, pH 7.0) transfer buffer and were run overnight. The blots were air-dried and UV cross-linked at 12,000 uJ/cm2 and stored at -20oC. They were screened with digoxigenin[PC1]-labeled probes at a concentration of 20 ng/ml of hybridization buffer.
characteristics.characteristic: description
characteristics.text: Matured third-instar larvae (diapausing and laboratory stored)
characteristics.characteristic: dev-stage
characteristics.text: Tetanops myopaeformis
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_454641
characteristics.characteristic: organism
characteristics.text: Differential display screening of dormant Tetanops myopaeformis
characteristics.characteristic: title
characteristics.text: plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pCR2.1
characteristics.characteristic: vector