|
| Status |
Public on Sep 29, 2010 |
| Title |
DEED+activin+RA (ExpI) |
| Sample type |
RNA |
| |
|
| Source name |
DEED+activin+RA (ExpI)
|
| Organism |
Xenopus laevis |
| Characteristics |
tissue: animal cap cells dissected from embryos injected with DEED antisense (DEED-As) nucleotide against Xenopus lhx1
treatment group: treated with activin and retinoic acid
|
| Biomaterial provider |
Neil Hukriede's lab
|
| Treatment protocol |
The embryos at two cell stage were injected with DEED-As nucleotide against Xenopus lhx1.The animal caps were dissected from the injected embryos at stage 9, and then treated with 1X10-4 M retinoic acid and 10ng/ml Activin.
|
| Growth protocol |
The DEED-As injected animal caps were cultured in 1X Danilchik’s solution with 1X10-4 M retinoic acid and 10ng/ml Activin till the sibling embryos reached stage 12.5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The dissected explants were homogenized in Stat 60 (TEL TEST), RNA was precipitated by isopropanol, treated with DNase I, and purified using the RNeasy kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated probe was prepared from 100 ng total RNA using the OVATION RNA amplification system (Nugen Technologies, Inc).
|
| |
|
| Hybridization protocol |
Total 2.2 microgram fargmented, biotin labeled amplified cDNA was used for hybridization, and the hybridization last for 18 hours.
|
| Scan protocol |
Scanning was performed by using GeneChip Scanner 3000 driven by GCOS1.4.
|
| Description |
Animal caps were injected with DEED-As, and then treated with activin and retinoic acid in experiment I.
|
| Data processing |
Single Array Analysis were performed by using GCOS1.4. Briefly, the “Target Signal” for “all probe set” was set to 500. Select “User Defined” and place a “1” as the "Normalization Value” in the “Normalization” tab. Detection p-value less than 0.04 is assigned a Present call, and above 0.06 is assigned an Absent call. Marginal calls are given to probe sets which have p-values between 0.04 and 0.06.
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| |
|
| Submission date |
Sep 28, 2010 |
| Last update date |
Sep 28, 2010 |
| Contact name |
Hui Zhao |
| E-mail(s) |
[email protected]
|
| Phone |
0085239431344
|
| Organization name |
The Chinese University of Hong Kong
|
| Department |
School of Biomedical Sciences
|
| Street address |
The Chinese University of Hong Kong
|
| City |
Hong Kong |
| ZIP/Postal code |
00000 |
| Country |
Hong Kong |
| |
|
| Platform ID |
GPL1318 |
| Series (1) |
| GSE24392 |
Lhx1 is required for specification of renal progenitor cell field |
|
