|
| Status |
Public on Sep 08, 2016 |
| Title |
Spiderbiofilm grown LR107 hht1 mutant (replicate 1) |
| Sample type |
RNA |
| |
|
| Source name |
Spider biofilm grown hht1 mutant
|
| Organism |
Candida albicans |
| Characteristics |
medium: Spider biofilm grown
genotype/variation: LR107 hht1 mutant
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated from C. albicans strains by the RNA Ambion kit (Cat. No. AM1560). For the biofilm induced cells, biofilm was developed in the Spider medium for 48 hours at 37°C
|
| 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 Whole Genome Candida albicans 8x15k (AMADID: 026377). 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 5190-0404). 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 |
Slides were scanned using the Agilent Microarray Scanner G2505C at 3 micron resolution.
|
| Description |
Spiderbiofilm grown LR107 hht1 mutant (replicate 1)
|
| Data processing |
Images were quantified using Feature Extraction Software ( Agilent). Feature extracted raw data was analyzed using GeneSpring GX software from Agilent. Normalization of the data was done in GeneSpring GX using the 75th percentile shift.
|
| |
|
| Submission date |
Sep 09, 2015 |
| Last update date |
Sep 08, 2016 |
| 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 |
GPL20624 |
| Series (1) |
| GSE72824 |
Regulation of the biofilm-specific genes circuitry in Candida albicans by a novel histone H3 variant |
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