|
| Status |
Public on May 01, 2015 |
| Title |
mature_hepatocyte_1 |
| Sample type |
RNA |
| |
|
| Source name |
RNA from mature human hepatocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cell: mature hepatocyte
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from foetal cells using TriZol (Invitrogen) according to the manufacturer's protocol. Total RNA from adult hepatocytes was purchased from 3H Biomedical, Uppsala, Sweden. The Nanodrop instrument (Labtech UK) was used to quantify RNA and assess 260/280 and 260/230 ratios. RNA integrity was examined using a 2100 Bioanalyzer (Agilent UK). All samples used had RIN scores > 9, 260/280 ratios > 2, and 260/230 ratios > 1.9.
|
| Label |
Hy3
|
| Label protocol |
1ug of total RNA was labelled with Hy3 dye according to the manufacturer's protocol. Briefly, RNA was incubated with the Hy3 dye, labelling enzyme, and spike-in miRNAs, in a total volume of 12.5ul for 1 hour at 16C. The enzyme was then heat-inactivated at 65C for 15 minutes. The samples were incubated at 95C for 2 minutes protected from light. Labelled samples were made up to 50ul by addition of 37.5ul of hybridization buffer.
|
| |
|
| Hybridization protocol |
The samples were briefly spun down and then loaded onto a MAUI hybridization station (BioMicro Inc., USA). The arrays were incubated at 56C for 16 hours, then washed briefly at 60C using buffer A, rinsed in buffer B, followed by a 2-minute wash in buffer B, a 2-minute wash in buffer C with a final brief fixing rinse in 99% ethanol. The arrays were dried by centrifugation (1000rpm for 5 minutes).
|
| Scan protocol |
Arrays were scanned using a GenePix 4200A microarray scanner.
|
| Data processing |
Raw scan data were analysed using GenePix Pro 6 software.
|
| |
|
| Submission date |
May 20, 2014 |
| Last update date |
May 01, 2015 |
| Contact name |
Iain Gallagher |
| E-mail(s) |
iaingallagher@gmail.com
|
| Phone |
00 44 1786 46 6024
|
| Organization name |
University of Stirling
|
| Department |
Faculty of Health Sciences and Sport
|
| Street address |
Room 4B133 Cottrell Building, University of Stirling, Airthrey Road
|
| City |
Stirling |
| ZIP/Postal code |
FK9 4LA |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL18712 |
| Series (2) |
| GSE57833 |
MicroRNA Expression profiles of mature hepatocytes and foetal liver progenitor cells |
| GSE57878 |
Profile of human liver progenitor cells |
|
