|
| Status |
Public on Apr 30, 2018 |
| Title |
PFNF54-Pfs16-GFP-LUC parasites 10 days after induction to produce gametocytes |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
PFNF54-Pfs16-GFP-LUC Day 10 RNA
|
| Organism |
Plasmodium falciparum |
| Characteristics |
strain: NF54-Pfs16-GFP-LUC
days after induction: 10
|
| Growth protocol |
P. falciparum NF54-Pfs16-GFP-LUC were induced by culturing in glucose-deficient media to produce gametocytes, before removing asexual parasites from the population by daily 5% D-sorbitol treatments from day 1-4 and thereafter culturing the parasites in glucose positive media and grown under stationary conditions for the entire time course.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples were harvested from culture by 0.01%(w/v) saponin addition on days -2 to 7 and via density centrifugation using Nycoprep 1.077 cushions on days 8-13 and stored at -80˚C until RNA isolation using TRIZOL reagent along with on-column extraction using the Qiagen RNeasy kit (Qiagen, Germany).
|
| Label |
Cy5
|
| Label protocol |
Painter, H. J., Altenhofen, L. M., Kafsack, B. F. C. & Llinás, M. Whole-genome analysis of Plasmodium spp. Utilizing a new agilent technologies DNA microarray platform. Methods Mol. Biol. 923, 213–219 (2013).
|
| |
|
| Channel 2 |
| Source name |
3D7 Mixed Stage asexual + NF54-Pfs16-GFP-LUC Mixed Stage gametocyte Reference Pool
|
| Organism |
Plasmodium falciparum |
| Characteristics |
strain: 3D7 + NF54-Pfs16-GFP-LUC
sample type: Reference Pool
|
| Growth protocol |
P. falciparum NF54-Pfs16-GFP-LUC were induced by culturing in glucose-deficient media to produce gametocytes, before removing asexual parasites from the population by daily 5% D-sorbitol treatments from day 1-4 and thereafter culturing the parasites in glucose positive media and grown under stationary conditions for the entire time course.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples were harvested from culture by 0.01%(w/v) saponin addition on days -2 to 7 and via density centrifugation using Nycoprep 1.077 cushions on days 8-13 and stored at -80˚C until RNA isolation using TRIZOL reagent along with on-column extraction using the Qiagen RNeasy kit (Qiagen, Germany).
|
| Label |
Cy3
|
| Label protocol |
Painter, H. J., Altenhofen, L. M., Kafsack, B. F. C. & Llinás, M. Whole-genome analysis of Plasmodium spp. Utilizing a new agilent technologies DNA microarray platform. Methods Mol. Biol. 923, 213–219 (2013).
|
| |
|
| |
| Hybridization protocol |
Painter, H. J., Altenhofen, L. M., Kafsack, B. F. C. & Llinás, M. Whole-genome analysis of Plasmodium spp. Utilizing a new agilent technologies DNA microarray platform. Methods Mol. Biol. 923, 213–219 (2013).
|
| Scan protocol |
Agilent G2600D Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version 11.5.1.1).
|
| Description |
Day 10.txt
|
| Data processing |
Agilent Feature Extraction Software (v 11.5.1.1) was used for background subtraction.
|
| |
|
| Submission date |
Oct 12, 2017 |
| Last update date |
Apr 30, 2018 |
| Contact name |
LynMarie Birkholtz |
| E-mail(s) |
lbirkholtz@up.ac.za
|
| Phone |
0124202479
|
| Organization name |
University of Pretoria
|
| Department |
Biochemistry
|
| Lab |
Malaria Parasite Molecular Laboratory
|
| Street address |
c/o Lunnon Road and Heroldt Street, Hatifeld
|
| City |
Pretoria |
| State/province |
Gauteng |
| ZIP/Postal code |
0083 |
| Country |
South Africa |
| |
|
| Platform ID |
GPL15130 |
| Series (1) |
| GSE104889 |
PFNF54-Pfs16-GFP-LUC gametocyte time course from commitment to maturity |
|
