LCK_1#37
Danio rerio
Protocols: The spleen from a heterozygote Tg(lck:EGFP) or wild-type fish was 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
2016-05-26
External Id
SAMEA4011512
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
2016-12-01T17:01:24Z
INSDC last update
2016-05-26T17:58:59Z
INSDC status
public
Submitter Id
E-MTAB-4617:LCK_1#37
broker name
ArrayExpress
common name
zebrafish
fsc
48296
gfp
195
individual
1
pi
4
plate
1
sample name
E-MTAB-4617:LCK_1#37
ssc
142
tissue
Spleen
well
E5