SAMN02299538
External Id
Developmental Genomics, LCDB, NIDDK, NIH
INSDC center name
2014-10-30T00:00:00Z
INSDC first public
2019-06-20T16:35:45.094Z
INSDC last update
SRS466574
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
SRS466574
SRA accession
100 ng
http://purl.obolibrary.org/obo/CHEBI_16412
amount of dna used to make library
S2
http://www.ebi.ac.uk/efo/EFO_0001233
cell line
anti-V5 (Invitrogen)
chip antibody
Drosophila melanogaster
http://purl.obolibrary.org/obo/NCBITaxon_7227
organism
S2 cells transfected with pMT5.1- DSXM-V5-His B
source_name
S2_DsxM
title
Transfection of S2 cells was performed using Effectene Transfection Reagent (Qiagen, Valencia, CA). pCoBlast (Invitrogen, Carlsbad, CA) was used as the selection plasmid. For the transfection procedure, cells were diluted to a density of 10^6 cells/ml and grew afterwards for about 20 hours. 1 µg expression plasmid, 50 ng pCoBlast, Effectene Transfection components and 840 µl complete Schneider’s Drosophila medium were mixed according to the manufacturer’s protocol and immediately added drop-wise onto the cells. The cells were subsequently grown in complete Schneider’s Drosophila medium for 60 hours prior to selection with 30 µg/ml blasticidin (Invitrogen, Carlsbad, CA). After 5 weeks of selection, the blasticidin-resistant cells were maintained in complete Schneider’s Drosophila medium containing 25 µg/ml of blasticidin. Expression of the recombinant proteins from the MT promoter was induced by adding copper sulfate to the medium to a final concentration of 500 μM. Before induction of expression cells were seeded to a density of 1*106 cells/ ml. Expression of the recombinant proteins was induced at a cell density of 3*10^6 cells/ ml. 60 ml of induced transfected Drosophila S2 cells expressing the recombinant proteins DSXM -V5-His or DSXF -V5-His respectively were grown in cell culture bottles for 36 hours reaching a cell density of 4.5*10^6 cells per ml.
treatment