|
| Status |
Public on May 03, 2016 |
| Title |
lame96_ area2_data |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
M+
|
| Organism |
Homo sapiens |
| Characteristics |
follicle size: M
related embryo development: +
|
| Treatment protocol |
freezing in liquid nitrogen until RNA extraction
|
| Growth protocol |
granulosa collected individually in vivo (ovum pick up)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
centrifugation on Qiashredder (Qiagen) 2min 13000 rpm, and total RNA extraction using phenol with miRNeasy kit (Qiagen) according to manufacturer guidlines.
|
| Label |
Cy5
|
| Label protocol |
Amplification protocol: This procedure describes the amplification of RNA using the RiboAmpb HSPlus RNA Amplification kit (Molecular devices). This procedure may be required for microarray experiments with extremely limited amounts of starting material. The RiboAmpb HSPlus RNA Amplification Kit enables the production of microgram quantities of antisense RNA (aRNA) from picogram quantities of total cellular RNA. The RiboAmpb HSPlus Kit achieves high yields of aRNA with a proprietary linear amplification process using double-stranded cDNA as template in a two round T7 RNA polymerase catalyzed amplification.
The labelling aRNA was done using the Universal Linkage System (ULS) by Kreatech for Gene Expression studies using microarray slides from Agilent. ULS is a non-enzymatic protocol that allows direct labeling of unmodified, amplified RNA. ULS will react with unmodified, amplified RNA and label it by formation of a coordinative bond on the N7 position of guanine. Fluorophore Cy3 and Cy5 are linked via a spacer to the ULS molecule.
|
| |
|
| Channel 2 |
| Source name |
S-
|
| Organism |
Homo sapiens |
| Characteristics |
follicle size: S
related embryo development: -
|
| Treatment protocol |
freezing in liquid nitrogen until RNA extraction
|
| Growth protocol |
granulosa collected individually in vivo (ovum pick up)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
centrifugation on Qiashredder (Qiagen) 2min 13000 rpm, and total RNA extraction using phenol with miRNeasy kit (Qiagen) according to manufacturer guidlines.
|
| Label |
Cy3
|
| Label protocol |
Amplification protocol: This procedure describes the amplification of RNA using the RiboAmpb HSPlus RNA Amplification kit (Molecular devices). This procedure may be required for microarray experiments with extremely limited amounts of starting material. The RiboAmpb HSPlus RNA Amplification Kit enables the production of microgram quantities of antisense RNA (aRNA) from picogram quantities of total cellular RNA. The RiboAmpb HSPlus Kit achieves high yields of aRNA with a proprietary linear amplification process using double-stranded cDNA as template in a two round T7 RNA polymerase catalyzed amplification.
The labelling aRNA was done using the Universal Linkage System (ULS) by Kreatech for Gene Expression studies using microarray slides from Agilent. ULS is a non-enzymatic protocol that allows direct labeling of unmodified, amplified RNA. ULS will react with unmodified, amplified RNA and label it by formation of a coordinative bond on the N7 position of guanine. Fluorophore Cy3 and Cy5 are linked via a spacer to the ULS molecule.
|
| |
|
| |
| Hybridization protocol |
The hybridization of microarray slides 4x44K printed with oligonucleotides by Agilent (Agilent-026652 human whole genome) using the expression array protocol.
|
| Scan protocol |
Scan on a PowerScanner (Tecan) using the autogain function. Images were quantified using ArrayPro version 6.3 (MediaCybernetics)
|
| Description |
Array 2
|
| Data processing |
Background subtracted, Loess and Quantile normalized data obtained from log2 of processed Red signal/processed Green signal. Flexarray 1.6.1 software used.
|
| |
|
| Submission date |
Dec 01, 2013 |
| Last update date |
May 03, 2016 |
| Contact name |
Anne-Laure Nivet |
| E-mail(s) |
anne-laure.nivet.1@ulaval.ca
|
| Organization name |
Laval University
|
| Street address |
2440 Hochelaga Blvd
|
| City |
Québec |
| State/province |
Québec |
| ZIP/Postal code |
G1V 0A6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL10332 |
| Series (1) |
| GSE52851 |
Follicular cells in vivo and developmental competence of oocytes in human |
|
