organism: Phallusia mammillata
description: Protocols: Adult P. mammillata animals were procured from the Roscoff Marine Station (France). P. mammillata embryos were dissociated with pronase for 20 minutes and individual dissociated embryos were transferred to calcium-free artificial sea water in imaging plates (Mat-tek) coated with a solution of 0.1%PFA and 0.1% gelatin. Dissociated embryos were kept on ice before collection. Individual cells were manually collected in 0.3 μl with pipet tips coated with a solution of 0.1%PFA and 0.1% gelatin and cells were directly lysed in 4μl of lysis+dNTP+oligodT mix (2 μl of lysis buffer – 0.2% Triton X-100+RNase inhibitor–, 1 μl of 10 μM oligo-dT primer – sequence 5'AAGCAGTGGTATCAACGCAGAGTACT30VN –, 1 μl of 10mM dNTP mix) according to the Smart-Seq2 protocol from Picelli et al. (2014). Cells were directly lysed in 4μl of lysis+dNTP+oligodT mix (2 μl of lysis buffer – 0.2% Triton X-100+RNase inhibitor–, 1 μl of 10 μM oligo-dT primer – sequence 5'AAGCAGTGGTATCAACGCAGAGTACT30VN –, 1 μl of 10mM dNTP mix) according to the Smart-Seq2 protocol from Picelli et al. (2014). cDNA from single cells was prepared following the Smart-Seq2 from Picelli et al. (2014). 500 pg of cDNA was then used for the tagmentation reaction using a home-made Tn5 transposase (Hennig et al., 2018).
attributes.tag: ENA first public
attributes.value: 2019-12-30
attributes.tag: ENA last update
attributes.value: 2018-02-21
attributes.tag: External Id
attributes.value: SAMEA104604944
attributes.tag: INSDC center alias
attributes.value: Cell Biology and Biophysics Unit. European Molecular Biology Laboratory
attributes.tag: INSDC center name
attributes.value: Cell Biology and Biophysics Unit. European Molecular Biology Laboratory
attributes.tag: INSDC first public
attributes.value: 2019-12-30T04:03:39Z
attributes.tag: INSDC last update
attributes.value: 2018-02-21T11:39:22Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6506:Sample 269
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: developmental stage
attributes.value: cleavage 16-cell
attributes.tag: individual
attributes.value: 16cell09097
attributes.tag: sample name
attributes.value: E-MTAB-6506:Sample 269