|
Status |
Public on Dec 01, 2010 |
Title |
Control replicate 2 at 48 hpf |
Sample type |
mixed |
|
|
Channel 1 |
Source name |
Total RNA from Zebrafish embroys, control treated
|
Organism |
Danio rerio |
Characteristics |
developmental stage: 48 hpf sample type: miRNAs and control RNAs from Zebrafish fish embroys tissue: embryo
|
Treatment protocol |
Embryos are collected from the mating cages and kept in an incubator at 28.5º C. Embroys are treated with 10nM Morphine sulphate , and 5hpto sacrifice time.
|
Growth protocol |
A natural breeding system has been used to obtain zebrafish embryos. This natural breeding is based on the photoperiod exhibited by the zebrafish in such a way that the egg laying is done during the first light of the day, so it can be controlled with artificial light.
|
Extracted molecule |
total RNA |
Extraction protocol |
Trizol® reagent (Invitrogen) was used to extract total RNA from cells
|
Label |
DY547
|
Label protocol |
Total RNA was ligated to a synthetic linker, 5’-pCU-DY547-3’. Reference DNA oligonucleotides (Invitrogen) complementary to a subset of mammalian miRNAs were combined and labeled with a ULYSIS® Alexa Fluor® 647 kit.
|
|
|
Channel 2 |
Source name |
Reference DNA
|
Organism |
Homo sapiens |
Characteristics |
sample type: synthetic DNA oligos corrresponding to human miRNAs
|
Treatment protocol |
Embryos are collected from the mating cages and kept in an incubator at 28.5º C. Embroys are treated with 10nM Morphine sulphate , and 5hpto sacrifice time.
|
Growth protocol |
A natural breeding system has been used to obtain zebrafish embryos. This natural breeding is based on the photoperiod exhibited by the zebrafish in such a way that the egg laying is done during the first light of the day, so it can be controlled with artificial light.
|
Extracted molecule |
other |
Extraction protocol |
Trizol® reagent (Invitrogen) was used to extract total RNA from cells
|
Label |
Alexa Fluor® 647
|
Label protocol |
Total RNA was ligated to a synthetic linker, 5’-pCU-DY547-3’. Reference DNA oligonucleotides (Invitrogen) complementary to a subset of mammalian miRNAs were combined and labeled with a ULYSIS® Alexa Fluor® 647 kit.
|
|
|
|
Hybridization protocol |
A mixture of labeled RNA and DNA was hybridized to a miRNA microarray (Thomson et al), inside a MAUI mixer in a MAUI hybridization system (BioMicro® System, Inc, Salt Lake City, UT, USA). Thomson, J.M., Parker, J., Perou, C.M., and S.M. Hammond. 2004. A custom microarray platform for analysis of microRNA gene expression. Nat. Methods 1: 47-53. Epub 2004 Sep 29.
|
Scan protocol |
Slides were scanned with a ScanArray 5000 machine (Perkin Elmer®, Waltham, MA, USA).
|
Description |
n/a
|
Data processing |
BlueFuse (BlueGenome, Cambridge, UK) was used to quantify pixel intensities. GeneSpring GX 7.3.1 (Agilent Technologies) was used for data normalization.
|
|
|
Submission date |
Nov 02, 2009 |
Last update date |
Jun 21, 2010 |
Contact name |
Xiaoxiao Zhang |
E-mail(s) |
zhang688@umn.edu
|
Organization name |
University of Minnesota
|
Street address |
312 Church St. SE
|
City |
Minneapolis |
ZIP/Postal code |
55455 |
Country |
USA |
|
|
Platform ID |
GPL9517 |
Series (1) |
GSE18847 |
microRNA expression and opioid regulation in the zebrafish |
|