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| Status |
Public on Mar 27, 2023 |
| Title |
HT29_METTL14 rep02 |
| Sample type |
RNA |
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| Source name |
HT29 human colon cancer cells, METTL14, replicate 2
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| Organism |
Homo sapiens |
| Characteristics |
genotype: p53 mutant
cell line: HT29 colorectal cancer cell
treatment: stably infected with lentivirus carrying METTL14 overexpression (METTL14)
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| Treatment protocol |
stably infected with control transfectants or stable METTL14 overexpression
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| Growth protocol |
HT29 human colon cancer cells were maintained in McCoy’s 5A supplemented with 10% fetal bovine serum (FBS) and antibiotics in humidified atmosphere with 5% CO2 at 37C
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| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from the HT29 human colon cancer cells by TRIzol reagent (Ambion) according to the manufacturer’s instructions. Globin message was further reduced using GLOBINclear (Ambion Inc., Austin, TX) to specifically remove both a- and b- globin. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
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| Label |
Cy3
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| Label protocol |
miRNA molecular in total RNA was labeled by miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) followed the manufacturer’s instructions, labeling section.
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| Hybridization protocol |
Each slide was hybridized with 100ng Cy3-labeled RNA using miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) in hybridization Oven (Cat # G2545A, Agilent technologies, Santa Clara, CA, US) at 55°C，20 rpm for 20 hours according to the manufacturer’s instructions, hybridization section. After hybridization, slides were washed in staining dishes (Cat # 121, Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit (Cat #5188-5327, Agilent technologies, Santa Clara, CA, US).
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| Scan protocol |
Slides were scanned by Agilent Microarray Scanner (Cat # G2565CA, Agilent technologies, Santa Clara, CA, US) and Feature Extraction software 11.5 (Agilent technologies, Santa Clara, CA, US) with default settings.
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| Data processing |
Raw data were normalized by Quantile algorithm, included in the R package AgiMicroRna(López-Romero, P. BMC Genomics. 2011.).
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| Submission date |
Jul 29, 2022 |
| Last update date |
Mar 27, 2023 |
| Contact name |
Yichao Hou |
| E-mail(s) |
[email protected]
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| Organization name |
Shanghai Jiao Tong University
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| Street address |
639 Zhi Zao Ju Road
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| City |
Shanghai |
| ZIP/Postal code |
200011 |
| Country |
China |
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| Platform ID |
GPL21575 |
| Series (2) |
| GSE210044 |
Gene expression profiling by miRNA microarray of control and METTL14-overexpressed HT29 human colon cancer cells |
| GSE210056 |
RNA-seq and miRNA microarray of control and METTL14-overexpressed HT29 human colon cancer cells |
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| Supplementary file |
Size |
Download |
File type/resource |
| GSM6415719_METTL14_02.txt.gz |
70.2 Kb |
(ftp)(http) |
TXT |
| GSM6415719_METTL14_02_raw_data.txt.gz |
7.0 Mb |
(ftp)(http) |
TXT |
| Processed data provided as supplementary file |
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