organism: Nematostella vectensis
description: Protocols: Animals were dissociated as described in Torres-Mendez et al., Nat Ecol Evol 2019. Cell suspensions were sorted by FACS to separate NvElav1::mOrange -positive from -negative cells. Total RNA was extracted with the Direct-zol RNA MicroPrep kit (Zymo Research). cDNA was prepared from 400pg of total RNA using the Smart-Seq 2 method with 16 pre-amplification PCR cycles, as described by Picelli et al. (2014). NGS libraries were prepared using the home-made tagmentation-based method as described by Hennig et al., 2018. Briefly, 125 ng of cDNA was tagmented using home-made Tn5 loaded with annealed linker oligonucleotides for 3 minutes at 55C. Reaction was inactivated by adding 1.25ul of 0.2% SDS and incubation for 5 minutes at room temperature. Indexing and amplification was done using the KAPA HiFi HotStart PCR kit (Sigma-Aldrich) with Index oligonucleotides (sequences were adapted from Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-02-28T04:03:31Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-02-13T12:43:35Z
attributes.tag: External Id
attributes.value: SAMEA6543562
attributes.tag: INSDC center name
attributes.value: Sars Centre for Marine Molecular Biology University of Bergen
attributes.tag: INSDC first public
attributes.value: 2020-02-28T04:03:31Z
attributes.tag: INSDC last update
attributes.value: 2020-02-13T12:43:35Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8794:NvElav::mOrange pos R4
attributes.tag: age
attributes.value: 14
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: starlet sea anemone
attributes.tag: developmental stage
attributes.value: primary polyp
attributes.tag: genotype
attributes.value: NvElav1::mOrange transgenics
attributes.tag: individual
attributes.value: pool of individuals
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: phenotype
attributes.value: NvElav1::mOrange positive
attributes.tag: sample name
attributes.value: E-MTAB-8794:NvElav::mOrange pos R4
attributes.tag: scientific_name
attributes.value: Nematostella vectensis
attributes.tag: sex
attributes.value: male and female
attributes.tag: strain
attributes.value: CH2 x CH6