GSM3756179: Knee-joint at E14.5 single-cell RNAseq by 10X Genomics; Mus musculus; RNA-Seq
Tissue was digested with 0.2% Collagenase II, 0.1% Dispase and 0.1% Hyaluronidase. Cells were washed twice with HBSS, and filtered through 40um cell strainer twice before loading onto the 10x genomics machine. Single cell encapsulation and cDNA libraries were prepared by Chromium™ Single Cell 3' Reagent Kits v2 and Chromium™ Single Cell A Chip Kit. Cells in suspension are counted and loaded into individual wells of 10X Chromium Single Cell chip. Single cells are then encapsulated into Gel in-beads emulsions (GEM) by 10X Chromium Controller. Single Cell 3' Reagent Kits v2 is used to perform downstream steps, in brief as follows: reverse transcription is performed on GEMs, followed by cDNA clean up and amplification. The double-stranded cDNA will then go through enzymatic fragmentation, adapter ligation, index PCR and SPRIselect size selection per manufacturer's protocol. Library size and concentration is determined by Qubit and Bioanalyzer assays. The libraries are denatured and diluted to optimal concentration. Cluster generation is performed on the flow cell with HiSeq PE Cluster Kit v4 by cbot. Illumina HiSeq SBS Kit v4 is used for sequencing run at 100 bp for read 1 and read 2.
GEO Accession
GSM3756179