|
| Status |
Public on Apr 11, 2010 |
| Title |
100Hz_LR_1h_DH5 rat |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
the tissue of spinal dorsal horn of 100Hz electroacupuncture low responder (LR) rats
|
| Organism |
Rattus norvegicus |
| Characteristics |
time point: 1h
strain: Sprague-Dawley
tissue type: spinal dorsal horn
frequency: 100Hz
group: low responder (LR) rats
|
| Treatment protocol |
Sprague-Dawley rats were received 100Hz electroacupuncture for 30 min, nociceptive testing and returned to home cages for 1 hours before sacrificed.These rats were non-sensitivity on EA analgesia. It's DH was then collected for transcript profiling analysis.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from DH using Trizol reagent (Invitrogen,USA), and further purified with a QIAGEN RNeasy Kit.
|
| Label |
Cy3
|
| Label protocol |
Total RNA were primed with T7 Promoter Primer (from the Agilent Low RNA Input Linear Amplification Kit PLUS, Two-Color) at 65C for 10 min, then reversed transcribed at 40C for 2 h in the presence of 1ul MMLV-RT (Agilent), and 10 mM dNTP, and RNase OUT (Agilent) and labeled with either Cy3-dCTP or Cy5-dCTP. Equal amounts of the RNA from control sample were mixed, and the mixture was labeled with Cy5, while the RNA from each DH which come from rats received 100Hz electroacupuncture for 30 min, nociceptive testing, returned to home cages for 1 hours and non-sensitivity on electroacupuncture analgesia was labeled with Cy3.
|
| |
|
| Channel 2 |
| Source name |
the tissue of DH of control rats
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague-Dawley
reference: rats sacrificed after stayed in their home cages without receiving electroacupuncture stimulation.
|
| Treatment protocol |
Sprague-Dawley rats were sacrificed after stayed in their home cages without receiving electroacupuncture stimulation.the RNA of DH were collected for transcript profilinf analysis
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from DH of control rats using Trizol reagent (Invitrogen,USA), and further purified with an QIAGEN RNeasy Kit
|
| Label |
Cy5
|
| Label protocol |
Total RNA were primed with T7 Promoter Primer (from the Agilent Low RNA Input Linear Amplification Kit PLUS, Two-Color) at 65C for 10 min, then reversed transcribed at 40C for 2 h in the presence of 1ul MMLV-RT (Agilent), and 10 mM dNTP, and RNase OUT (Agilent) and labeled with either Cy3-dCTP or Cy5-dCTP. Equal amounts of the RNA from control sample were mixed, and the mixture was labeled with Cy5, while the RNA from each DH which come from rats received 100Hz electroacupuncture for 30 min, nociceptive testing, returned to home cages for 1 hours and non-sensitivity on electroacupuncture analgesia was labeled with Cy3.
|
| |
|
| |
| Hybridization protocol |
After mixed with hybridization buffer(GE healthcare), samples were applied to microarray slide, cover the slip with care. Place the slide in a wet box.Incubate in a hybridization chamber at 42°C for 16-18 hours, avoid light exposure. After hybridization, slides were washed sequentially with 1 SSC/0.2 SDS(50°C), 0.1 SSC/0.2%SDS(50°C) and 0.1 SSC(room temperature) before scanning.
|
| Scan protocol |
Axon GenePix 4000B
|
| Description |
Total RNA was prepared by using TRIzol reagent (Life Technologies, USA), further purified with an RNEasy column (Qiagen, Germany), and quantified by using gel electrophoresis and spectrophotometer followed by reverse transcription labeling. Each RNA sample was reverse-transcribed into cDNA primed with oligo(dT) and labeled with either Cy3-dCTP or Cy5-dCTP by using Superscript II reverse transcriptase (Life Technologies, USA). After hybridization under a standard protocol, data acquisition was performed by using a laser scanner (Agilent, USA), and the image data were analyzed using ImaGene 4.2 (Biodiscovery, Santa Monica, CA, USA).
|
| Data processing |
LOWESS normalized, background subtracted VALUE data obtained from log of processed Red signal/processed Green signal.
|
| |
|
| Submission date |
Apr 08, 2010 |
| Last update date |
Dec 14, 2010 |
| Contact name |
ke wang |
| E-mail(s) |
[email protected]
|
| Phone |
+86 21 51320288
|
| Organization name |
Shanghai Biochip Co., Ltd.
|
| Street address |
151 Libing Road
|
| City |
Shanghai |
| ZIP/Postal code |
201203 |
| Country |
China |
| |
|
| Platform ID |
GPL3498 |
| Series (2) |
| GSE21258 |
Transcript Profiling of Spinal Dorsal Horn in Response to Electroacupuncture on Rats at 1h |
| GSE21758 |
Transcript Profiling of Spinal Dorsal Horn in Response to Electroacupuncture on Rats at two time points |
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