15 protocols
AccessionType
bioassay_data_transformation
ID_REF = VALUE = Processed signals intensity with Feature Extraction Software (not normalized).
image_aquisition
Hybridization signals were detected with a DNA microarray scanner G2505A (Agilent Technologies).
feature_extraction
The scanned images were analyzed by using Agilent feature extraction software (v9.5.1.1).
hybridization
Labeled RNA was hybridized to Human Whole Genome Microarray kit (Agilent) at 65 degrees C for 24hr rotating at 10 rpm and washed with Gene Expression Hybridization Kit (Agilent) according to the manufacturer protocol (Version 5.7).
image_aquisition
Hybridization signals were detected with a DNA microarray scanner G2505B (Agilent Technologies).
feature_extraction
The scanned images were analyzed by using Agilent feature extraction software (v9.1.3.1).
hybridization
Labeled RNA was hybridized to microarray at 60 degrees C for 17hr and washed with miRCURY microarray labeling kit (Exiqon) according to the manufacturer protocol.
labeling
Total RNA (0.5 µg) was labeled and hybridized by using Quick Amp Labeling Kit (Agilent Technologies) according to the manufacturer protocol (Version 5.7).
labeling
Total RNA (2 µg) was labeled with miRCURY microarray labeling kit (Exiqon) according to the manufacturer protocol.
specified_biomaterial_action
For miRNA trasnfection, myxoid liposarcoma-derived cells were trasnfected with 250 nM in DMEM without serum using Oligofectamine Reagent (Invitrogen Japan, Tokyo, Japan) and Opti-MEM I (Invitrogen Japan).
grow
The myxoid liposarcoma-derived cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS).
nucleic_acid_extraction
Total RNA was extracted from liposarcoma cell-line with ISOGEN (NIPPONGENE) following the manufacture's protocol.
grow
NIH3T3 cells were cultivated in D-MEM (Life technologies) supplemented with 10% fetal calf serum (Gibco BRL, Rockville, MD).
specified_biomaterial_action
NIH3T3 (2.2×105) cells were seeded in 6 cm plates. 16 hours later, they were transfected by 5 μg of DNA (TLS/CHOP type 2 vector and control vector) according to the FuGENER HD Transfection protocol (Qiagen, Tokyo, Japan).
nucleic_acid_extraction
Total RNA was extracted with ISOGEN (NIPPONGENE) following the manufacture's protocol.