|
Status |
Public on Mar 04, 2020 |
Title |
B.subtilis_t = 60_rep1 |
Sample type |
RNA |
|
|
Source name |
Bacillus subtilis wild type
|
Organism |
Bacillus subtilis |
Characteristics |
strain: PY79 development phase: phase dark spore
|
Growth protocol |
Purified spores were heat activated (HA) (70°C for 30 min) prior to germination. The spores were suspended at 4.5* 10˄10 spores/ml in MOPS liquid minimal medium (pH 7.4) supplemented with a mixture of AGFK (10mM L-asparagine, 10mM D-glucose, 1mM D-fructose, 1mM KCl) and 10mM L-alanine. Throughout the spore revival, samples were drawn at regular time intervals for further analysis
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using the RNeasy MinElute Cleanup kit from Qiagen and DNA was removed by Turbo DNA-free kit (Ambion Inc., Applied Biosystems, USA). The purity and quantity of isolated RNA were checked spectrophotometrically on a NanoDrop spectrophotometer (ND-1000, Isogen Life Science, NL).
|
Label |
Cy3
|
Label protocol |
Labeling was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
|
|
|
Hybridization protocol |
Each hybridization mixture comprised 1.1 μg Test (Cy3) and 1.1 μg Reference (Cy5), a common reference pool of all samples. Microarrays were with the NimbleGen Hybridization System (Roche NimbleGen).
|
Scan protocol |
Scanning was performed by NimbleScan v2.6 (Roche NimbleGen).
|
Data processing |
For the raw data Log2 ratios were calculated for each feature and gene expression values were calculated using the median polish algorithm
|
|
|
Submission date |
Mar 03, 2020 |
Last update date |
Mar 04, 2020 |
Contact name |
Bhagyashree Swarge |
E-mail(s) |
b.n.swarge@uva.nl
|
Organization name |
Swammerdam institute of life sciences, University of Amsterdam
|
Street address |
Science park 908
|
City |
Amsterdam |
State/province |
North Holland |
ZIP/Postal code |
1098XH |
Country |
Netherlands |
|
|
Platform ID |
GPL16974 |
Series (1) |
GSE146277 |
Intergrative analysis of proteome and transcriptome dynamics during Bacillus subtilis spore revival |
|