|
| Status |
Public on Dec 31, 2016 |
| Title |
Hemocytes of infected larvae of B. mori 1 |
| Sample type |
RNA |
| |
|
| Source name |
Hemocytes of infected larvae of B. mori
|
| Organism |
Bombyx mori |
| Characteristics |
tissue: Hemocyte
sample type: Infested
|
| Treatment protocol |
Haemolymph was collected at 6h after invasion of E. bombycis maggot, and hemocytes were pelleted by centrifugation and used for RNA extraction
|
| Growth protocol |
B. mori larvae were infested with E. bombycis on day 1 fifth instar.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA isolation was done by using QIAGEN RNeasy mini kit (Cat#74106) as per the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
The samples were labeled using Agilent Quick-Amp labeling kit (Part number: 5190-0424). 500ng of total RNA was reverse transcribed using oligo dT based method. mRNA was primed with oligo dT primer tagged to T7 promoter sequence and converted into double stranded cDNA using MMLV-RT reverse transcriptase. Further, in the same reaction cDNA was in-vitro transcribed to cRNA using T7 RNA polymerase enzyme. During cRNA synthesis, Cy3 labeled Cytosine nucleotide was incorporated into the newly synthesized strands. Labeled cRNA thus obtained was cleaned up using Qiagen RNeasy columns (Qiagen, Cat No: 74106). Concentration and amount of dye incorporated were determined using Nanodrop.
|
| |
|
| Hybridization protocol |
1650ng of labeled cRNA were hybridized on the array (AMADID: 66047) using the Agilent Gene Expression Hybridization kit (Part Number 5190-0404) in Sure hybridization Chambers (Agilent) at 65ºC for 16 hours. Hybridized slides were washed using wash buffers (Part No: 5188-5327; Agilent).
|
| Scan protocol |
Scanned using the Agilent Microarray Scanner (Agilent Technologies, Part Number G2600D) at 5 micron resolution.
|
| Description |
Hemocytes of infected larvae of B. mori
|
| Data processing |
Images were quantified using Feature Extraction Software ( Agilent).
|
| |
|
| Submission date |
Nov 30, 2015 |
| Last update date |
Dec 31, 2016 |
| Contact name |
Genotypic technology |
| E-mail(s) |
sudha.rao@genotypic.co.in
|
| 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 |
GPL20728 |
| Series (1) |
| GSE75520 |
Host- parasite interaction: Transcriptome responses to parasitism in the silkworm Bombyx mori |
|
