|
| Status |
Public on Oct 20, 2011 |
| Title |
Nimotuzumab 3 |
| Sample type |
RNA |
| |
|
| Source name |
Nimotuzumab
|
| Organism |
Homo sapiens |
| Characteristics |
source cell line: A431
tissue: tumor xenograft
treatment: Nimotuzumab
|
| Treatment protocol |
Six doses of each treatment were given to the mice intra peritonealy. Dosings were given on day 7,9,11,14,16 and 18. The mice were then sacrificed on Day 18, two hours after the last dosing.
|
| Growth protocol |
A431 cells were grown in DMEM medium (10% FBS), 5*10^6 cells were injected in to the mice sub cutaneously.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tissue was homogenized using Tissue Homogenizer and RNA isolation was done using TRIzol method.
|
| Label |
Cy3
|
| Label protocol |
The samples for Gene expression were labeled using Agilent Quick-Amp labeling Kit (p/n5190-0442). Five hundred nanograms each of the Control and test samples were incubated with reverse trancription mix at 40°C and converted to double stranded cDNA primed by oligodT with a T7 polymerase promoter. Synthesized double stranded cDNA were used as template for cRNA generation. cRNA was generated by in vitro transcription and the dye Cy3 CTP(Agilent) was incorporated during this step. The cDNA synthesis and in vitro transcription steps were carried out at 40°C. Labeled cRNA was cleaned up and quality assessed for yields and specific activity.
|
| |
|
| Hybridization protocol |
The labeled cRNA samples were hybridized on to a Genotypic designed Custom Whole Genome Human 8x60k (AMADID No: 027114). 600ng of cy3 labeled samples were fragmented and hybridized. Fragmentation of labeled cRNA and hybridization were done using the Gene Expression Hybridization kit of Agilent (Part Number 5188-5242). Hybridization was carried out in Agilent’s Surehyb Chambers at 65º C for 16 hours. The hybridized slides were washed using Agilent Gene Expression wash buffers (Part No: 5188-5327)
|
| Scan protocol |
The labeled cRNA samples were hybridized on to a Genotypic designed Custom Whole Genome Human 8x60k (AMADID No: 027114). 600ng of cy3 labeled samples were fragmented and hybridized. Fragmentation of labeled cRNA and hybridization were done using the Gene Expression Hybridization kit of Agilent (Part Number 5188-5242). Hybridization was carried out in Agilent’s Surehyb Chambers at 65º C for 16 hours. The hybridized slides were washed using Agilent Gene Expression wash buffers (Part No: 5188-5327)
|
| Description |
Gene data after tumor extracted on Day 18
|
| Data processing |
Data extraction from Images was done using Feature Extraction software Version 10.7 and Normalization of the data was done in GeneSpring GX V 11.5
|
| |
|
| Submission date |
Sep 23, 2011 |
| Last update date |
Oct 20, 2011 |
| Contact name |
Genotypic technology |
| E-mail(s) |
[email protected]
|
| Organization name |
Genotypic Technology
|
| Street address |
259, Apoorva 4th cross,80 feet Road,RMV 2ND STAGE
|
| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560094 |
| Country |
India |
| |
|
| Platform ID |
GPL13252 |
| Series (1) |
| GSE32333 |
Xenograft mouse study in C57BL6/SCID mice using A431 cells |
|
