Project description:This metabolomics dataset was generated on an Exploris 480, and contains Pellet and Supernatant sample fractions of various Algae samples from the Bigelow Marine Labs.
Project description:This metabolomics dataset was generated on an Exploris 480, and contains Pellet and Supernatant sample fractions of various Algae samples from the Bigelow Marine Labs.
Project description:HEK293T cells were cultured according to standard protocols and heat shocked at 42oC for 6 hours. Detergent-soluble and insoluble fractions from HEK293T cells were obtained following the procedures described before. Specifically, HEK293T cells were lysed in NP40 lysis buffer (Invitrogen #FNN0021) with protease inhibitors by gentle pipetting. The cell homogenate was centrifuged at 14000 rpm and the supernatant was retrieved (detergent-soluble fraction). The remaining pellet was washed twice in ice-cold NP40 lysis buffer and then resuspended in urea buffer (RIPA buffer with 8M urea). The samples were then centrifuged at 14000 rpm for 10 min to pellet any remaining cell debris. The resulting supernatant (detergent-insoluble fraction) was retrieved and analyzed as explained below.
Project description:Circulating miRNAs are an emerging class of biomarkers correlating their specific expression patterns to disease states. A considerable proportion of hematopoietically-derived miRNAs are present in plasma with the ability to confound the signature of true circulating miRNA species. We use microarray analysis to catalogue a list of 313 haemotopoetic miRNAs and analyze expression profiles of cell-free miRNAs in individual plasma fractions after calibrating for cellular miRNA signals. Comprehensive global maps of bona fide circulating miRNA species are presented, and inter-individual variability and gender-specific expression is explored in populations of healthy individuals. Healthy Caucasian individuals were used to segregate blood into 8 subfractions: white blood cells (WBC)/buffy coat, leukocytes, red blood cells (red blood cells (RBC)), cloudy supernatant (CS), supernatant 1 (S1), supernatant 2 (S2), pellet 1 (P1) and pellet 2 (P2) through differential centrifugation to understand the proportion of cellular miRNAs in each category.
Project description:Crosslinked DNA from wild type cells or from mutant of the THO complex was analyzed by tiling arrays to precisely detect DNA targets of THO in yeast. DNA coming from SDS-washed DCF pellet and the supernatant SN2k of 2 strains were compared in the study, WT (W303) and mft1 delta (DLY224). The DNA of each condition (Pellet or Supernatant) was labeled either with Cy3 or Cy5 fluorescent dyes and competitively hybridized on 244k agilent arrays. For each strain, the experiment was performed on 2 biological replicates, with dye swap.
Project description:200,000 worms were grown to the young adult stage, collected, and washed. Worms were homogenized in lysis buffer (10mM Tris, pH 7.5, 138mM NaCl, 2mM CaCl2, and 0.1% NP-40). After homogenization, 125U/mL Benzonase nuclease was added to homogenates and samples were incubated for 30 mins at room temperature with gentle rocking. Samples were spun at 21,000g for 5 mins, followed by resuspension of the pellet in lysis buffer. Soluble proteins were removed from the pellet with repeated washes in lysis buffer until A280 of the supernatant was equal to A280 of lysis buffer. Resulting pellet was resuspended in ddH2O, spun at 21,000g for 5 mins, and supernatant was transferred to a collection tube. Water resuspensions and recoveries were continued until A280 of H2O supernatants was 0. Recovered water washes were pooled and amyloids salted out by adding NaCl to 200mM final concentration, followed by 1-3 days incubation at 4oC. Precipitated amyloids were recovered by centrifuging at 21,000g for 30 mins, followed by resuspension in 50-100uL ddH2O. Amyloids were digested (trypsin and chymotrypsin) and prepared for mass spectrometry as described in the methods files.
Project description:Reticulocytes were purified from fresh blood samples obtained from 10 healthy adult volunteers (5 women and 5 men) after informed consent. The donors had normal blood cell counts, blood smears, hemoglobin electrophoresis, red cells membrane resistance tests and no biological evidence of hemolysis. Whole blood samples were centrifuged, the supernatant and the buffy coat containing white blood cells (WBC) and platelets were removed. The red cells pellet was then purified using the method described by Brun et al. Purity, assessed by Hematology Flow Cytometer was 1 leukocyte per 15 millions RBC and efficiency of purification process was 99.8% and 1 platelet per 10 000 RBC. Keywords: other
Project description:Proteomic data from pig ligated loops. Pig loops infected with C. jejuni F38011 and mutant; time points 3, 12, 30 hours post infection. Supernatant and pellet samples collected from loops; 6 biological replicates for supernatant samples and 5 biological replicates for pellet samples.
Project description:Clavibacter michiganensis subsp. michiganensis is an important Gram-positive phytopathogenic bacteria that causes bacterial wilt and canker in tomato. The genome of the type strain, NCPPB382, has been sequenced and annotated, however comparative genomics suggests that certain regions are under- or misannotated. In order to improve the genome annotation, we have undertaken a proteogenomic study of this important pathogen. Samples were grown in culture and the proteome of the pellet and supernatant were analyzed separately using shotgun HPLC-MS/MS. These proteomics datasets were analyzed and a number of missing gene were found and a number of existing gene calls were modified.