Vibrio cholerae Freezing Study
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ABSTRACT:
ORGANISM(S): Vibrio cholerae
PROVIDER: GSE38179 | GEO | 2013/05/23
SECONDARY ACCESSION(S): PRJNA167440
REPOSITORIES: GEO
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Project description:Question Addressed: Does gene expression in V. cholerae change when samples are frozen at -80 degrees? An in vitro grown culture of O1 Inaba ICDDR,B (strain DSM-V999 described in Nature. 2002 Jun 6;417(6889):642-5) that had been grown to exponential phase (OD600 + 0.2) was placed in a 15 ml conical and then placed in a -80 degree freezer. A portion of the starting culture was harvested prior to freezing to serve as the control/reference for downstream hybridizations. RNA was recovered from the non-frozen sample as well as by prepping material directly from frozen samples. Labeling reactions were performed in quadruplicate for each sample. A replicate experimental design type is where a series of replicates are performed to evaluate reproducibility or as a pilot study to determine the appropriate number of replicates for a subsequent experiments. replicate_design
2013-05-23 | E-GEOD-38179 | biostudies-arrayexpress
Project description:This portion of the study evaluated the effects of topical 10% black raspberry gel application on gene expression profiles of premalignant oral lesions. Topical application of the bioadhesive black rasberry gel was observed to modulate gene expression and reduce proinflammatory proteins in human premalignant oral lesions. Experiment Overall Design: Each patient served as their own internal control. A pretreatment specimen of the premalignant lesion was obtained for baseline parameters. Following six weeks of treatment, the treated lesional tissue was removed. All tissue samples were immediately placed in RNA later and placed in a -80 degree freezer until sample preparation.
2008-06-18 | E-GEOD-10174 | biostudies-arrayexpress
Project description:These experiments were performed to show serogroup conversion in Vibrio cholerae from O1 to O139 in a mixed communities / biofilms. For this purpose, V. cholerae O1 El Tor A1552 and VCO139-Kan strain (a MO10 derivative; O139 serogroup) were grown on crab shell fragments to induce natural competence for transformation. Transformants were selected on LB+Kan+Rif plates. O139 positive transformants have undergone a full exchange of the O1 region by the O139 region. This implies an exchange of an at least 32 kb spanning O1 genomic region by more than 42 kb of the O139 region. The transformation experiment was done at least five independent times; data from four experiments are shown; per experiment one to three clones were analysed by CGH with two experimental replicates each. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Keywords: all_pairs, array CGH
2007-04-25 | GSE5932 | GEO
Project description:Transcriptional profiling to investigate the effect of drug treatment on the E. faecalis cells. For microarray analysis, E. faecalis OG1RF was grown in FMC medium supplemented with 10 mM glucose to an optical density at 600 nm (OD600) of 0.3 and the cultures were divided in 7 aliquots. One aliquot was collected by centrifugation and immediately frozen (untreated control cells). The other aliquots were treated for 30 or 60 min with 1.25 X the minimum inhibitory concentration (MIC) of one of the following CW inhibitors: ampicillin (20 μg ml-1), bacitracin (80 μg ml-1), or cephalothin (80 μg ml-1). After an exposure time of 30 or 60 minutes, each of these cultures was also centrifuged and the pellets frozen. RNA was then isolated from each pellet for microarray analysis. This process was repeated 3 additional times, for a total of four replicates of each condition.
2013-03-16 | GSE45231 | GEO
Project description:GSM306248: S. cerevisiae was grown on YEPD. Wild type and msg5 yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 4hours. After 4 hours of growth cells were collected, frozen at -80 °C and processed for RNA extraction. GSM306249: msg5 yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 4hours. After 4 hours the culture was divided into two parts. One was treated with alpha factor and the other part was allow to grow without any treatment. After 90 minutes of treatment, both cultures of cells were collected, frozen at -80 °C and processed for RNA extraction.
2008-12-31 | GSE12104 | GEO
Project description:ngs2015_12_zeawall2-zeawall2-Address the hypothesis that allelic variation at a major QTL for cell wall related traits gives rise to coordinated changes in gene expression in maize-Two pairs of BC2S2 maize lines carrying F271 or F288 alleles in an 18-20 Mb interval of chromosome 6 in an otherwise F271 genetic background were grown in the field at Mauguio (Hérault, France) in 2015 with two row replicates. Irrigation was applied during the summer to prevent water stress. The bottom third of upstream-ear internodes (without nodes) of three representative plants in each of the two replicates were harvested two days before silking. All samples were immediately frozen in liquid nitrogen and stored at -80°C.
2019-12-14 | GSE115157 | GEO
Project description:Gene expression was examined during four growth conditions which modulated antimicrobial secretion in B. ambifaria AMMD. A minimal salts medium (BSM; Hareland et al. 1975. J Bacteriol 121:272-85) with 0.05% yeast extract was used for all comparisons with the carbon source (4 g/L) and growth condition varied as follows: (i) liquid culture with glucose; (ii) liquid culture with glycerol; (iii) growth on agar with glucose, and (iv) growth on agar with glycerol. For the liquid cultures, 25 ml of growth medium was placed a 250 ml conical flask and inoculated with 0.5 ml of a starter culture of strain AMMD which had been optically adjusted to O.D. 1 at 600 nm and corresponded to a viable count of 10 million colony forming units per ml. After growth with shaking (150 rpm) for 30 h at 30 degrees Celcius, the O.D. 600 nm of the cultures was measured to enable an estimation of cell density to be made, they were then snap-chilled and harvested as previously described (Drevinek et al. 2008. BMC Infect Dis 8:121), and frozen at -80 degrees Celcius. Growth on agar was performed by laying a sterile 47 mm nitrocellulose filter (0.22 ?m) onto a plate of the BSM agar and the same starter culture of AMMD as used for the liquid growth spread over a 2 cm diameter circular area using a sterile cotton swab. The agar cultures were incubated for 30 h at 30 degrees Celcius, the filters removed to sterile 50 ml conical tubes and the bacteria washed off by gentle pipeting using 2 ml ice cold BSM (without any carbon source or yeast extract). This resuspension of surface grown bacteria was processed prior to RNA extraction in an exactly the same way as the liquid cultures
2011-07-13 | E-MEXP-2819 | biostudies-arrayexpress
Project description:These experiments were performed to show a serogroup conversion in Vibrio cholerae from O1 to O139. For this purpose, V. cholerae O1 WT = A1552 was grown on crab shell fragments to induce natural competence for transformation. Purified DNA (2 ug each) from strain VC73-orf6/7-Kan-A was added after 24h and the cells grown further for 24h. The VC73-orf6/7-Kan-A strain is a ATCC25873 derivative (both O37 serogroup) which harbors a Kanamycin cassette in the O37 region (as part of the operon between orf6 and orf7 w/o own promotor) for better selection. Transformants were selected on LB+Kan plates. Three clones were selected from each experiment and analyzed by microarray hybridization (BioPrime. Array CGH Genomic Labeling from Invitrogen). Two microarray replicates were done per clone. Comparison of A1552 versus VC73-orf6/7-Kan-A is shown as control. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Keywords: genotyping_design
2007-04-26 | GSE5935 | GEO
Project description:These experiment were performed to show a serogroup conversion in Vibrio cholerae from O1 to O139. For this purpose, V. cholerae O1 WT = A1552 was grown on crab shell fragments to induce natural competence for transformation. Purified DNA (2 ug each) from strain VCO139-Kan was added after 24h and the cells grwon further for 24h. The VCO139-Kan strain is a MO10 derivative (both O139 serogroup) which harbors a Kanamycin cassette in the O139 region (as part of the operon between wbfA and wbfB w/o own promotor) for better selection. Transformants were selected on LB+Kan plates. Two groups of transformants were gained: Group I had a full exchange of the O1 region by the O139 region (clones serogroup-converted: SGC#1-3); the crossovers for the homologous recombination event had occurred within or upstream of the gmhD gene and in most instances within or downstream of the homolog gene of VC0271. This implies an exchange of an at least 33 kb spanning O1 genomic region by more than 42 kb of the O139 region. Group II had only half of the O139 region transfered and therefore half of the O1 region kept (clones HSGC#4-6). We analyzed their genotype and found that all of them had undergone a homologous recombination event with one crossover in or upstream of the gmhD gene and the second one inside the VC0254 and IS1358 gene. The transformation experiment was done three independent times (I - III). Three clones from group I and group II were selected from each experiment and analyzed by microarray hybridization (BioPrime. Array CGH Genomic Labeling from Invitrogen). Two microarray replicates were done per clone. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Strain Name: -III for experimental replicates (transformation); clones #1 -#3 full region exchanged; clones #4-#6 half region exchanged. Two MA replicates for each clone Keywords: genotyping_design, array CGH
2007-04-25 | GSE5120 | GEO
Project description:These experiments were performed to show a serogroup conversion of Vibrio cholerae from O1 to 37. For this purpose, V. cholerae O1 El Tor (A1552) was grown on crab shell fragments to induce natural competence for transformation. Purified DNA (4 ug each) from strain ATCC25872, an O37 serogroup strain, was added after 24h and the cells were further grown for 24h. After detachment from the crab shell fragments, bacteria were poured into soft-agar and overlaid onto LB plates. Mukerjee's El Tor phage III (a gift of Dr. M.S. Islam) was dropped onto the surface of the bacteria containing soft-agar. The plaques formed by killing non-transformed A1552 cells possessed resistant clones which were picked and further selected by non-agglutination with O1-specific antiserum. One to four clones were selected from each independent experiment and analyzed by microarray hybridization (BioPrime. Array CGH Genomic Labeling from Invitrogen). Two microarray replicates were done per clone. Strain Names: AIpO37#1 / AIpO37#4 / AIpO37#6 / AIpO37#8 are clones analyzed from the first experiment; AIIpO37#9 / AIIpO37#13 / AIIpO37#16 are clones analyzed from the second independent experiment; AIIIpO37#9A is a clone analyzed from the third independent experiment; AIVpO37#1A / AIVpO37#3A / AIVpO37#4A / AIVpO37#8A are clones analyzed from the fourth independent experiment. Two MA replicates per clone were done. CGHs of A1552 versus ATCC25872 are provided as control. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Keywords: genotyping_design
2007-04-25 | GSE5934 | GEO