Protein and Nucleic Acid Analysis Core
Calvin Vary, PhD
Protein, Nucleic Acid Analysis and Cell Imaging Core at MMCRI has been established to offer high quality, reproducible sequences and characterization of peptides and nucleic acids.
Automated DNA sequencing is offered on both shorter and longer instruments depending on the sample. DNA and RNA extraction is offered by robotic extraction and purification. High throughput (16-capillary) sequencing is available for appropriate projects. Deep sequencing/NextGen sequencing is available via interaction with the Tufts University Core facilities-so please inquire.
Protein sequencing and post-translational analysis is offered using Mass Spectrometry by means of protein digestion and subsequent peptide fingerprinting and compositional analysis. This facility houses nanoscale Liquid Chromatography interfaced to Quadrapole Mass Spectrometer (LC/MS) for protein identification, quantitative differential analysis using isotope-tagging approaches (e.g. ICAT), Post-translational modification analysis and protein-protein interaction identification can also be accomplished. Informatics analysis uses state-of-art automated (e.g. ProteinPilot) determination or post translational modification, chemical adduction (ICAT, ITrac) , phosphorylation, gel-based modification etc.
Under the auspices of the National Childrens' Study, we have recently added a linear ion trap 4000QTRAP mass spectrometer and its dedicated RSLC_3000 nanoscale liquid chromatograph. In addition to protein identification, this instrument is capable if isotope-free quantitative analysis of peptides or small molecules using the technique of multiple reaction monitoring (MRM).
Recent example of Mass spectrometry applications:
Romero D, O'Neill C, Terzic A, Contois L, Young K, Conley BA, Bergan RC, Brooks PC, Vary CPH. Endoglin regulates cancer-stromal cell interactions in prostate tumors. Cancer Res. 2011:15;71(10):3482-93.
Oligonucleotide and siRNA ordering through IDT
DNA Sequencing: (Service Request Form)
- DNA sequencing (run only)
- DNA sequencing (core does cleanup and run)
HPLC, QSTAR and 4000QTRAP: (Service Request Form)
- Nano-liquid chromatography
- Protein identification
- Protein post translational modification
- Peptide digestion, fingerprinting and sequencing
- Multiple reaction monitoring (MRM)
Service forms must be filled out completely with as much detailed information as possible. The completed form must include the signature and account number of the Principle Investigator of the lab requesting services. Completed forms should be submitted to Barbara Conley (Vary Lab).
This component of the Protein, Nucleic Acid Analysis and Cell Imaging Core is supported by the Vascular Biology Center of Biomedical Research Excellence (COBRE) Program of the National Center for Research Resources.