|
| Status |
Public on Jun 22, 2015 |
| Title |
Day5-6 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Day5-rep3
|
| Organism |
Sus scrofa |
| Characteristics |
developmental stage: Day5 blastocyst
|
| Extracted molecule |
total RNA |
| Extraction protocol |
This document applies to the total RNA extraction using the PicoPure RNA Isolation Kit from Molecular Devices. The PicoPure RNA Isolation Kit enables to recover total cellular RNA from pico-scale samples. Researchers can obtain high recoveries of total cellular RNA from as little as a single cell to samples with up to 100 ug of RNA. RNA extraction from larger samples should be done using another kit or technique.
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.
|
| Label |
Cy5
|
| Label protocol |
This document describes the procedure used for labelling aRNA 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 |
Day5-rep2
|
| Organism |
Sus scrofa |
| Characteristics |
developmental stage: Day5 blastocyst
|
| Extracted molecule |
total RNA |
| Extraction protocol |
This document applies to the total RNA extraction using the PicoPure RNA Isolation Kit from Molecular Devices. The PicoPure RNA Isolation Kit enables to recover total cellular RNA from pico-scale samples. Researchers can obtain high recoveries of total cellular RNA from as little as a single cell to samples with up to 100 ug of RNA. RNA extraction from larger samples should be done using another kit or technique.
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.
|
| Label |
Cy3
|
| Label protocol |
This document describes the procedure used for labelling aRNA 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 |
80 μl of hybridization mixture containing 8 ul 20X SSC, 3.2 ul 2% SDS, 4.8 ul Liquid Blocking reagent (Amersham, USA), 120pmol of samples labelled with each dye
|
| Scan protocol |
GenePix 4200 Autoloader by Molecular Devices. Software is GenePix Pro, Acuity 4.0
|
| Data processing |
background subtracted, data obtained from log2 of processed Red signal/processed Green signal
|
| |
|
| Submission date |
Jun 25, 2012 |
| Last update date |
Jun 22, 2015 |
| Contact name |
chi zhou |
| Organization name |
University of Alberta
|
| Department |
Agriculture Food and Nutritional Science
|
| Street address |
114 street 90 ave NW.
|
| City |
Edmonton |
| State/province |
Alberta |
| ZIP/Postal code |
T6G 2P5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL7435 |
| Series (1) |
| GSE38923 |
Day5-6 porcine blastocyst gene expression profile |
|
