Project description:This study evaluates a resequencing microarray designed to determine the sequence of the four major genes in the Ebolavirus genome, Nucleoprotein (NP), matrix protein (VP40), glycoprotein (GP) and polymerase (L). The array has the ability to determine the sequence of five species and strains: Zaire Ebolavirus (Mayinga and Makona), Bundibugyo Ebolavirus, Sudan Ebolavirus and Tai Forest Ebolavirus. Illumina Next Generation Sequencing verified the sequence of the Zaire Ebolavirus (Mayinga) sample.
Project description:Bisulfite conversion and whole genome-single base next generation sequencing of DNA from a single iPSC clone (CMC28). This method provides exceptional depth of the sequenced methylome. Bisulfite converted DNA from a single iPSC clone (CMC28), and get its high-throughput sequence data with Illumina.
Project description:This study demonstrates the ability of an Ebolavirus resequencing microarray to determine the sequence of the Zaire ebolavirus glycoprotein that has been engineered into the place of the surface protein of a recombinant vesicular stomatitis virus expressing green fluorescent protein (rVSV-EBOVgp-GFP). The rVSV-EBOVgp-GFP was cultured in VERO-E6 cells for three passages either in the presence of a monoclonal antibody that blocks infection (KZ52) or in control cultures with no antibody. Culture supernatant and cell lysate was collected before passaging and after each passage. RNA was extracted from each sample and the sequence of the Ebola glycoprotein in each sample was determined by the Ebola resequencing microarray. Illumina Next Generation Sequencing was performed on the initial virus stock, before passaging and on the third passage of the KZ52 antibody selected virus stock to validate the microarray sequence results.
Project description:GATA4 occupancy on the mouse genome of satellite cell-derived primary myoblasts. Proliferating myoblasts cultured in growth medium were immunoprecipitated with anti-GATA4 antibody or control IgG. Precipitated genomic DNAs were subjected to next generation sequencing. Paired-end 150 bp sequence reads of GATA4-ChIP and IgG-ChIP using mouse skeletal muscle myoblasts.
Project description:A custom resequencing array for analysis of field isolates of plasmdium falciparum was created. Test of DNA with genotypes known at all loci genotyped by the microarray as well as test of accuracy correlation with amounts of DNA added to each array Comparison of test DNA from lab and clinical isolates between genotyped generated by next-generation sequencing and this new custom DNA microarray.
Project description:The twelve zinc finger Suppressor of Hairy-wing [Su(Hw)] protein binds thousands of sites in Drosophila genome and is essential for the function of the gypsy insulator. Loss of the globally expressed Su(Hw) protein causes female sterility due to tissue-specific defect limited to female germline. Using chromatin immunoprecipitation followed by next generation sequencing (ChIP-Seq), we determine the extent of tissue-specific binding of Su(Hw) in Drosophila ovary. We demonstrate that Su(Hw) binding sites (SBSs) are largely constitutively occupied in the germline and soma. Our analyses indicate that SBSs fall into several non-uniform classes, as determined by the partner protein distribution and DNA sequence conservation. Further, we show that only a subset of SBSs is required for the female fertility. These sites are maintained in the su(Hw)f zinc finger 10 mutant background, which is fertile but does not support gypsy insulator function. Together, our data are consistent with the model where Su(Hw) serves multiple regulatory roles in the genome, and contribute to understanding of how loss of a single zinc finger affects chromosome localization of a DNA binding protein. Examination of Su(Hw) localization in the ovaries of less than 6 hour old wild type and su(Hw)f mutant Drosophila females
Project description:Investigation of whole genome gene expression level changes in a Azospirillum lipoferum 4B associated to artificial roots, Oryza sativa japonica cv. Cigalon roots and Oryza sativa japonica cv. Nipponbare roots, compared to the strain grown in liquid culture. For each of the four condition, two replicates were analysed on an A. lipoferum 4B whole genome expression array designed by Roche Nimblegen, Inc. (Madison, WI, USA), based on the genome sequence (Wisniewski-DyM-CM-) et al. 2011), as follows: two replicates of 5 probes (length, 60 nucleotides) per gene, covering 6,242 genes and using a total of 62,178 probes.
Project description:One of the cornerstones of an effective biodefense strategy is the ability to detect infectious agents with a high degree of sensitivity and specificity in the context of a complex sample background. The nature of the B. anthracis genome, however, renders specific detection difficult, due to close homology with B. cereus and B. thuringiensis. We therefore elected to determine the efficacy of next-generation sequencing analysis and microarrays for detection of B. anthracis in an environmental background. We applied next-generation sequencing to titrated genome copy numbers of B. anthracis in the presence of background nucleic acid extracted from aerosol and soil samples. We found next-generation sequencing to be capable of detecting as few as 10 genomic equivalents of B. anthracis DNA per nanogram of background nucleic acid. Detection was accomplished by mapping reads to either a defined subset of reference genomes or to the full GenBank database. Moreover, sequence data obtained from B. anthracis could be reliably distinguished from sequence data mapping to either B. cereus or B. thuringiensis. We also demonstrated the efficacy of a microbial census microarray in detecting B. anthracis in the same samples, representing a cost-effective and high-throughput approach, complementary to next-generation sequencing. This Series contains the NimbleGen array data only (no next-generation sequencing data). B. anthracis DNA was spiked at 6 different concentrations (1, 10, 100, 1000, 10000 and 100000 genome copies) into 1 ng of background nucleic acids extracted either from a soil sample or from an aerosol (air filter) sample. Two replicates of each combination of B. anthracis copy number and background sample were analyzed.
Project description:With the whole genome SNPs array information, we could evaluate the sensitivity and specificity of the point mutation we conclude from the next-generation sequencing data. Furthermore, we could use the true positive mutation as our guidance to exclude the most unreliable single nucleotide variation detected from sequence. After the process, we could promise a very high specificity under minimum loss of sensitivity. To evaluate the sensitivity and specificity of point mutaions detected through the next-generation sequencing data from nine cases of M5 leukemia patients.