A Ph-positive chronic myelogenous leukemia cell line treated with Imatinib.
Treatment protocol
5 microMol of imatinib for 12 hrs
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using a RNeasy Mini Kit (Qiagen, Germantown, MD, USA).The amount of RNA was measured by NanoDrop (NanoDrop Technologies, Wilmington, DE, USA), then the quality of extracted RNA was checked using a 2100 Bioanalyzer (Agilent Technologies, Wilmington, DE, USA).
Label
Cy5
Label protocol
20 microgram of RNA were labeled using a LabelStar Array kit (Qiagen) , according to the supplier’s instruction.
Total RNA was extracted using a RNeasy Mini Kit (Qiagen, Germantown, MD, USA).The amount of RNA was measured by NanoDrop (NanoDrop Technologies, Wilmington, DE, USA), then the quality of extracted RNA was checked using a 2100 Bioanalyzer (Agilent Technologies, Wilmington, DE, USA).
Label
Cy3
Label protocol
20 microgram of RNA were labeled using a LabelStar Array kit (Qiagen) , according to the supplier’s instruction.
Hybridization protocol
Hybridization was carried out automatically using GeneTAC Hybstation (Genomic Solutions, US) according to the supplier’s instruction. The conditions of hybridization were 55oC for 2 hours, 50oC for 2 hours, and 46oC for 2 hours, then 42oC for 12 hours.
Scan protocol
The hybridized signals were scanned by GenePix 4000B Microarray Scanner (Axon Instruments, Union City, CA, USA).
Description
Imatinib (STI571; Novartis, Basel, Switzerland), a potent and selective inhibitor of the tyrosine kinase activity of BCR-ABL
Data processing
The scanned data was normalized, verified, and analyzed using the Genomic Profiler software (Mitsui Industrial Knowledge, Inc, Tokyo, Japan:http://bio.mki.co.jp/en/product/index.html).
