|Public on Oct 01, 2023
|mouse embryonic cells
|tissue: aorta-gonads-mesonephros (AGM)
cell type: AEC
developmental stage: E10.0
|FACS-sorted cells were kept on ice until lysed and reverse transcribed.
|Wildtype AECs and HECs were sorted from the E10 embryonic AGM region of C57 mice.
|Morphologically deformed cells were discarded. Living single cells were rinsed in PBS-BSA, and manually transferred into cell lysis buffer with a mouth pipette; 0.05 μl of 1:200,000 dilution of External RNA Controls Consortium (ERCC) RNA spike-in Mix1 (Ambion) was added to lysis buffer per reaction.
The cDNA fragments were end-repaired and added dA-tailed using Ultra II End Prep module and then tethered to 1D adaptor by using Quick Ligation Module. After that, each cDNA library was loaded into 1 FLOPRO002 flow cell and sequenced on PromethION.Beta.
PolyA enriched single-cell Nanopore Sequencing
|Guppy (V3.5.2) software used for basecalling.
Sequenced reads were firstly processed with nanopack, then mapped to GENCODE (vM22) mouse genome (mm10) with minimap2 (v2.18-r1015) with default parameters
Isofrorm-level expression was quantified as counts with Salmon (version 1.1.0).
Supplementary_files_format_and_content: Tab-delimited text files include RPT10K at isoform level.
|Oct 08, 2021
|Last update date
|Oct 01, 2023
|Peking Union Medical College
|Dong Dan San Tiao 5, Dongcheng District
|Single-cell Architecture and Functional Requirement of Alternative Splicing during Hematopoietic Stem Cell Emergence [RNA-Seq]
|Single-cell Architecture and Functional Requirement of Alternative Splicing during Hematopoietic Stem Cell Emergence