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Status |
Public on Jan 01, 2016 |
Title |
SNP cells |
Sample type |
RNA |
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Source name |
Canine mammary gland tumor
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Organism |
Canis lupus familiaris |
Characteristics |
gender: female age: 10-year-old tissue: tumor cells in the pleural effusion
|
Treatment protocol |
Establishment of the cell line.The tumor cells were prepared under aseptic conditions for cell culture. The cells were cultured in 35-mm diameter petri dishes (Nunc, Ltd., Roskilde, Denmark) in RPMI 1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% heat-inactivated fetal bovine serum (Nichirei Biosciences Inc., Tokyo, Japan) and PSN (5 mg/ml penicillin, 5 mg/ml streptomycin, and 10 mg/ml neomycin) solution (Invitrogen), then incubated in 5% CO2 at 37 °C. The cells were subcultured by washing with phosphate-buffered saline. Then, the cells were harvested from near-confluent cultures by brief exposure to a solution containing 0.25% trypsin and 1 mmol/l tetrasodium ethylenediaminetetraacetic acid solution with phenol red (Invitrogen). Trypsinization was stopped using RPMI 1640 containing 10% fetal bovine serum. Trypsinized cells were transferred to a new petri dish.
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Growth protocol |
For clonal experiments, we serially diluted cell suspensions and plated them onto 96-well plates. We marked wells containing one single cell after microscopic confirmation. A colony was then sub-cultured from each marked well into a 12-well plate. Cloning was repeated two times. Cell lines were maintained in continuous culture over 60 passages, and we designated the established cell line as SNP cells.
|
Extracted molecule |
total RNA |
Extraction protocol |
miRNeasy® Mini Kit was used.
|
Label |
biotin
|
Label protocol |
We followed protocol of manual that is FlashTagTM Biotin HSR RNA Labeling Kit User Manual.
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|
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Hybridization protocol |
We followed protocol of manual that is FlashTagTM Biotin HSR RNA Labeling Kit User Manual.
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Scan protocol |
We followed manufacturer's protocol, and used GeneChip® Scanner 3000 7G.
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Data processing |
Data acquisition and normalization (RMA) were used Affymetrix® Expression Console™ Software.
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Submission date |
Mar 20, 2015 |
Last update date |
Jan 01, 2016 |
Contact name |
Tomohiro Osaki |
Organization name |
Tottori University
|
Department |
Faculty of Agriculture
|
Street address |
4-101 Koyamachominami
|
City |
Tottori |
State/province |
Tottori |
ZIP/Postal code |
680-8553 |
Country |
Japan |
|
|
Platform ID |
GPL19117 |
Series (1) |
GSE67129 |
Establishment and miRNA expression characterization of a canine mammary gland tumor cell line |
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