LCK_8#65
Danio rerio
Protocols: Spleens from two heterozygote Tg(lck:EGFP) fish were dissected and passed through a 40μm cell strainer using the plunger of a 1-mL syringe and cells were collected in cold 1xPBS/5% FBS. A non-transgenic line was used to set up the gating and exclude autofluorescent cells. Propidium iodide (PI) staining was used to exclude dead cells. Individual cells were sorted, using a Becton Dickinson Influx sorter with 488- and 561 nm lasers (Schulte et al., 2015) and collected in a single well of a 96 well plate containing 2.3 uL of 0.2 % Triton X-100 supplemented with 1 U/uL SUPERase In RNAse inhibitor (Ambion). The size, granularity and level of fluorescence for each cell were simultaneously recorded. The Smart-seq2 protocol (Picelli et al., 2014) was used to amplify the whole transcriptome and prepare libraries. Twenty-five cycles of PCR amplification were performed.
ENA first public
2016-12-01
ENA last update
2016-09-12
External Id
SAMEA4432053
INSDC center alias
Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland Swiss Institute of Bioinformatics
INSDC center name
Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland Swiss Institute of Bioinformatics
INSDC first public
2016-12-01T17:01:25Z
INSDC last update
2016-09-12T14:14:52Z
INSDC status
public
PI fluorescence
57
Submitter Id
E-MTAB-46171473686057:LCK_8#65
age
3
broker name
ArrayExpress
common name
zebrafish
developmental stage
adult
forward scatter
32758
genotype
heterozygote Tg(lck:EGFP)
gfp fluorescence
1034
individual
2
organism part
spleen
plate
8
sample name
E-MTAB-46171473686057:LCK_8#65
side scatter
44
single cell well quality
good quality
well
F5
well information
single cell