|
| Status |
Public on Apr 14, 2010 |
| Title |
Small intestine-juvenile (NF66) Replicate-2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Dissected intestine from ~5 juvenile frogs (NF66)
|
| Organism |
Xenopus laevis |
| Characteristics |
developmental stage: Juvenile frogs (NF61)
tissue: Small intestine
|
| Growth protocol |
X. laevis tadpoles were maintained and used in accordance with the guidelines established by NICHD Animal Use and Care Committee. They were sacrificed and inteestines were collected at the indicated developmental stages according to the same established guidelines.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using TRIZOL following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
2 µg of total RNA were used for each sample for cDNA synthesis and subsequent labeled Cy3-labeled cRNA synthesis using Low Input Labeling Kit (two-color) (Agilent Technologies) following manufacturer's instructions. Universal Reference cRNA (labeled with Cy5) was prepared from 200 ng of mRNA isolated from pooled mRNA for multiple stages representing all tissues and all the metamorphic stages of Xenopus laevis using the same Low Input Labeling Kit (two-color) (Agilent Technologies) following manufacturer's instructions .
|
| |
|
| Channel 2 |
| Source name |
Universal Reference mRNA made from pooled mRNA for multiple stages representing all the metamorphic stages of Xenopus laevis.
|
| Organism |
Xenopus laevis |
| Characteristics |
developmental stage: All stages from NF50 to NF66
tissue: All tissues
|
| Growth protocol |
X. laevis tadpoles were maintained and used in accordance with the guidelines established by NICHD Animal Use and Care Committee. They were sacrificed and inteestines were collected at the indicated developmental stages according to the same established guidelines.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using TRIZOL following manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
2 µg of total RNA were used for each sample for cDNA synthesis and subsequent labeled Cy3-labeled cRNA synthesis using Low Input Labeling Kit (two-color) (Agilent Technologies) following manufacturer's instructions. Universal Reference cRNA (labeled with Cy5) was prepared from 200 ng of mRNA isolated from pooled mRNA for multiple stages representing all tissues and all the metamorphic stages of Xenopus laevis using the same Low Input Labeling Kit (two-color) (Agilent Technologies) following manufacturer's instructions .
|
| |
|
| |
| Hybridization protocol |
Agilent 60-mer oligo microarray processing protocol (Agilent_2_color_Microarray-based Gene Exp Analysis.pdf, version 5.5). The manual can be found at Agilent support website or through a WEB search.
|
| Scan protocol |
Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1).
|
| Description |
Biological replicate 2 of 3. Dissected small intestine from juvenile frog (NF66).
|
| Data processing |
Agilent Feature Extraction Software (v 8.5.1.1) was used for data extraction from scanner generated TIFF files. Local background subtraction and LOWESS normalization settings were used.The data were further processed with NIA ANOVA tool utilities.See http://lgsun.grc.nia.nih.gov/ANOVA for details.
|
| |
|
| Submission date |
Apr 12, 2010 |
| Last update date |
Apr 13, 2010 |
| Contact name |
Biswajit Das |
| E-mail(s) |
dasbiswa@mail.nih.gov
|
| Organization name |
National Institutes of Health/National Institute of Child Health and Human Development
|
| Department |
Laboratory of Gene Regulation and Development
|
| Lab |
Section on Molecular Morphogenesis
|
| Street address |
18 Library Drive, Building 18T, Room 106
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL10302 |
| Series (1) |
| GSE21303 |
Studies on Xenopus laevis intestine reveal biological pathways underlying vertebrate gut adaptation from embryo to adult |
|
