|
| Status |
Public on Oct 01, 2015 |
| Title |
AP2-G2(-)_19h_4 |
| Sample type |
RNA |
| |
|
| Source name |
AP2-G2(-)_19h
|
| Organism |
Plasmodium berghei ANKA |
| Characteristics |
genotype/variation: AP2-G2 KO
infection host: rat
tissue: blood synchronously infected with P. berghei parasites
time point: 19 h after the innoculation
|
| Growth protocol |
Whole blood was harvested from rats infected with AP2-G2(-) or wild-type parasites and cultured for 16 h. Mature schizonts were purified from the culture with Nicodenz and inoculated into rats. Whole blood was harvested from these rats at 19 h after the innoculation. Four biologically independent experiments were performed in both AP2-G2(−) and wild-type parasite samples.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocols.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 1.0 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.65 ug Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scannedon the Agilent DNA Microarray Scanner (G2565BA) using one color scan setting for 1x44k array slide.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters.
|
| |
|
| Submission date |
Feb 23, 2015 |
| Last update date |
Oct 01, 2015 |
| Contact name |
masao yuda |
| E-mail(s) |
m-yuda@doc.medic.mie-u.ac.jp
|
| Organization name |
mie university
|
| Street address |
edobashi 2-174
|
| City |
tsu |
| State/province |
mie |
| ZIP/Postal code |
514-0001 |
| Country |
Japan |
| |
|
| Platform ID |
GPL19816 |
| Series (2) |
| GSE66188 |
Global Transcriptional Repression: Initial and Essential Step for Plasmodium Sexual Development [expression array] |
| GSE66190 |
Global Transcriptional Repression: Initial and Essential Step for Plasmodium Sexual Development |
|
