|
| Status |
Public on Nov 29, 2013 |
| Title |
DIA3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
diapausing mite_whole organism
|
| Organism |
Tetranychus urticae |
| Characteristics |
developmental stage: adult
gender: female
|
| Treatment protocol |
By letting mites develop in diapause inducing conditions, some mites entered into diapause, while others did not.
|
| Growth protocol |
Larvae of the LS-VL strain were kept in diapause inducing conditions (17±0.5 °C, 80% RH and 8:16h light:dark photoperiod) until reaching the adult female life stage.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from adult female spider mites using RNEasy Mini Kit (Qiagen) following manufacturer's instructions.
|
| Label |
cy5
|
| Label protocol |
100 ng of total RNA (+ RNA-spike in control) was used to generate Cy3 or Cy5 labeled cRNAs, using the Agilent Low Input Quick Amp Labeling Kit (version 6.5 Agilent Technologies)
|
| |
|
| Channel 2 |
| Source name |
non-diapausing mite_whole organism
|
| Organism |
Tetranychus urticae |
| Characteristics |
developmental stage: adult
gender: female
|
| Treatment protocol |
By letting mites develop in diapause inducing conditions, some mites entered into diapause, while others did not.
|
| Growth protocol |
Larvae of the LS-VL strain were kept in diapause inducing conditions (17±0.5 °C, 80% RH and 8:16h light:dark photoperiod) until reaching the adult female life stage.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from adult female spider mites using RNEasy Mini Kit (Qiagen) following manufacturer's instructions.
|
| Label |
cy3
|
| Label protocol |
100 ng of total RNA (+ RNA-spike in control) was used to generate Cy3 or Cy5 labeled cRNAs, using the Agilent Low Input Quick Amp Labeling Kit (version 6.5 Agilent Technologies)
|
| |
|
| |
| Hybridization protocol |
Cy3 and Cy5 labeled cRNA were pooled and hybridized using the Gene Expression Hybridization Kit (Agilent Technologies) for 17h in a rotating hybridization oven at 20 r.p.m. and 65°C; Slides were washed using the Gene Expression Wash Buffer Kit (also provided by Agilent Technologies)
|
| Scan protocol |
Hybridized arrays were scanned using an Agilent Microarray High Resolution Scanner.
|
| Description |
Replicate 3 of 4, DIA vs NON-DIA
Sample 3
|
| Data processing |
Data were normalized by the Agilent Feature Extraction software, using the GE2_107_Sep09 protocol. This protocol uses a Linear and Lowess algorithm for normalization. See also page 35 in the reference guide of the Agilent Feature extraction software (http://www.chem.agilent.com/Library/usermanuals/Public/G4460-90026_FE_Reference.pdf).
|
| |
|
| Submission date |
Jul 15, 2013 |
| Last update date |
Nov 30, 2013 |
| Contact name |
Nicky Wybouw |
| Organization name |
University of Ghent
|
| Department |
Department of Crop Protection
|
| Street address |
Coupure Links 653
|
| City |
Gent |
| ZIP/Postal code |
9000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL16890 |
| Series (1) |
| GSE48858 |
Genome wide gene-expression analysis of facultative reproductive diapause in the two-spotted spider mite Tetranychus urticae |
|
