| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Single cell suspension was prepared as previously described with some modifications. Briefly, planarians were finely diced on ice plate and washed off the plate with CMFB buffer, followed by digestion with 1% trypsin in CMFB (CMF+0.5%BSA) for 8 min at room temperature with gentle agitation, and then pipetting up and down the suspension to break up the clumps. After strained through a 40 mm filter, cells were pelleted by centrifugation at 290 g for 5 minutes at 4°C and stained with Hoechst 33342 (Thermo, 62249) for 30 minutes at room temperature. Before loading onto MoFlo cell sorter, cells were strained again and then Propidium iodide (Thermo, J66584.AB) was added to discriminate dead cells. For single-cell RNA-seq from 10x Chromium platform, Hoechst-stained and PI negative cells (200,000 cells) from wild-type animals were collected on ice using cell sorter. The libraries were made using the Chromium platform and Chromium Single Cell 3’ v3 chemistry. Sequencing libraries were loaded on an Illumina nova 6000 flowcell with two 150bp paired-end kits. |
RNA-Seq |
TRANSCRIPTOMIC |
unspecified |
PAIRED
|
|
