|
| Status |
Public on Jul 30, 2009 |
| Title |
0.2 fmole of miRNAs |
| Sample type |
mixed |
| |
|
| Channel 1 |
| Source name |
miRNAs from mirVana miRNA Reference Panel v9.1
|
| Organism |
synthetic construct |
| Characteristics |
synthetic RNAs coresponding to human miRNAs
|
| Extracted molecule |
total RNA |
| Extraction protocol |
commercially available
|
| Label |
DY547
|
| Label protocol |
Different amounts of miRNA standards in the panel were ligated to a synthetic linker 5’-pCU-DY547-3’. 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 |
synthetic
|
| Organism |
synthetic construct |
| Characteristics |
synthetic DNA oligos corrresponding to human miRNAs
|
| Extracted molecule |
other |
| Extraction protocol |
commercially available
|
| Label |
Alexa Fluor® 647
|
| Label protocol |
Different amounts of miRNA standards in the panel were ligated to a synthetic linker 5’-pCU-DY547-3’. 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 |
We hybridized different amounts of microRNAs from the mirVana miRNA Reference Panel v9.1 to microarrays.
|
| 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 |
Apr 15, 2008 |
| Last update date |
Jan 20, 2009 |
| 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 |
GPL6743 |
| Series (1) |
|
