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Status |
Public on Dec 06, 2010 |
Title |
E2-xGem injected st.10.5 |
Sample type |
RNA |
|
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Source name |
embryonic ectoderm
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Organism |
Xenopus laevis |
Characteristics |
developmental stage: st. 10.5 manipulation: xGem injected
|
Treatment protocol |
xGemininΔcoil RNA/300 pg was injected into both blastomeres of two-cell stage pigmented Xenopus laevis embryos (animal hemisphere). Injected embryos were raised until stage 8-9 and 80 ectodermal (animal cap) explants/sample were isolated and incubated in 0.7X MMR/25ºC.
|
Growth protocol |
Xenopus laevis embryos were obtained by in vitro fertilization
|
Extracted molecule |
total RNA |
Extraction protocol |
When sibling embryos reached stage 10.5 or 12, total RNA (20 μg/sample) was isolated using Trizol (Invitrogen).
|
Label |
biotin
|
Label protocol |
biotin
|
|
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Hybridization protocol |
Performed by the Washington University Genome Technololgy Access Center
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Scan protocol |
Affymetrix Genechip 3000 used by the Washington University Genome Technololgy Access Center
|
Description |
xGem st.10.5-set 2.CEL
|
Data processing |
To define microarray target genes, raw CEL and DAT files were analyzed with dChip software (http://biosun1.harvard.edu/complab/dchip/)(Li and Hung Wong, 2001) after normalization to exclude probesets that did not meet the following preliminary cutoff values: i) expression, represented by model-based expression indices (MBEI), was changed more than 2-fold in treated versus untreated samples (E/B >2.0) groups, ii) MBEI differences were larger than 70 (E-B > 70), iii) expression was claimed as “present (P)” in at least one sample in each experiment. Probesets were considered putatively Geminin-regulated if they met microarray cutoff values in at least 2 of the 3 experiments. A subset of these genes was then further validated.
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Submission date |
Nov 05, 2010 |
Last update date |
Dec 06, 2010 |
Contact name |
Kristen L Kroll |
E-mail(s) |
kkroll@wustl.edu
|
Organization name |
Washington University School of Medicine
|
Department |
Developmental Biology
|
Lab |
Kristen Kroll
|
Street address |
320 McDonnell Sciences/660 S. Euclid Ave.
|
City |
Saint Louis |
State/province |
MO |
ZIP/Postal code |
63110 |
Country |
USA |
|
|
Platform ID |
GPL1318 |
Series (1) |
GSE25158 |
Geminin-regulated genes in the Xenopus laevis embryonic ectoderm |
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