title: MBhold2K_library_1
organism: Macrotermes bellicosus
description: Protocols: Heads were flushed with RNAlater™ (Qiagen) to ensure complete permeation of the head tissue. The samples were transported on ice to the field station and kept overnight at 4 °C to ensure full permeation. Then samples were kept frozen in a -20 °C fridge not more than two weeks. During transport, the samples were kept in an insulated container, cooled on ice, until arrival in the laboratory in Freiburg (Germany). There, samples were kept in a -80 °C freezer until further processing. Total RNA was extracted from the head of single individuals according to a modified version of the peqGOLD TriFast™ protocol: modifications were an incubation time of 10 min instead of 5 min at room temperature after adding 900 μl peqGOLD Tri Fast™ and inverting the tube. After adding chloroform and after incubation, centrifugation was modified to 10 min at 4 °C, 10,800 rpm. After transfer of the aqueous phase into a new Eppendorf tube, we added another centrifugation step for 5 min at 4 °C (10,800 rpm). Then the supernatant was transferred into a new Eppendorf tube and 2 μl nuclease free glycogen (5 mg/ml) was added per sample. After adding isopropanol, the mixture was incubated for 10 min at room temperature and not on ice. After washing the RNA pellet with 75% ethanol and vortexing, the sample was centrifuged for 5 min at 4 °C, 8,500 rpm. This step was repeated three times. After dissolving the pellet in nuclease-free water, samples were kept for at least one hour at 4 °C. DNA digestion was performed using DNase I Amplification Grade Kit (Sigma-Aldrich). The concentration and quality of the isolated RNA was checked with an Agilent Bioanalyzer (Agilent RNA 6000 Nano Kit). Library preparation was done by BGI. RnaseH treatment was performed additionally to the Ribo-Zero® rRNA Removal Kit (Human/Mouse/Rat), Illumina (http://www.illumina.com/products/ribo-zero-rrna-removal-human-mouse-rat.html).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-04-26T17:04:17Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-04-23T15:37:20Z
attributes.tag: External Id
attributes.value: SAMEA4603151
attributes.tag: INSDC center name
attributes.value: University of Freiburg Institute for Biology I (Zoology) Evolutionary Biology & Ecology
attributes.tag: INSDC first public
attributes.value: 2018-04-26T17:04:17Z
attributes.tag: INSDC last update
attributes.value: 2018-04-23T15:37:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6767:MBhold2K_library_1
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: organism part
attributes.value: head
attributes.tag: phenotype
attributes.value: minor worker
attributes.tag: sample name
attributes.value: E-MTAB-6767:MBhold2K_library_1
attributes.tag: sampling site
attributes.value: Africa, Cote d'Ivoire, Comoe National Park, Coordinates 8.6550568, -3.7823524 +- 1 km, 200m
attributes.tag: scientific_name
attributes.value: Macrotermes bellicosus