FAQ

SPARC BioCentre gives priority to strategic collaborative research projects based at SickKids but services may be made available other members of the U of T community and beyond after consultation. 

There are numerous precautions that can be taken to ensure that there is minimal keratin contamination as well as chemical interference with your samples.  The following is only a guide; however, care should be taken at all steps of the sample preparation process.  This is perhaps the most important part of the process!

Wear gloves and lab coats at all times. Furthermore, use clean reagents, glassware, and plasticware to avoid keratin contamination from your skin and dust ie. new bags of tips and microcentrifuge tubes.

Specific to SDS-PAGE Gels:

• If you are running a SDS-PAGE gel ahead of sample preparation, please consider using pre-cast gels, or at very minimum use new glass plates, as well as clean, new, boxes for staining.
• When cutting gel bands out, use clean materials and work in a laminar flow hood whenever possible. Cut bands using a clean scalpel, and cut out each protein band.  Gel slices should not be wider then 2-3 mm. Cutting exactly around the band, avoiding excess gel, drastically improves your results.
• Do not combine multiple bands into the same tube
• Gel slices should be stored in 1% acetic acid at 4^oC prior to analysis.

Specific to in-solution samples:

• Detergents (NP-40, Triton-X, Tween-20) can lead to the detection of a polymer-like background on the mass spectrometer, and can bind to the C18 column, subsequently preventing your peptides from binding. Avoid these detergents if possible; however, if they are necessary, we can employ techniques to remove them.
• While SDS does not produce a polymer like background, significant amounts of SDS are also not compatible with mass spectrometry. Again, we can employ techniques to remove SDS at certain concentration levels, but be sure to let us know if your samples contain SDS.
• In the case of immunoprecipitation samples, detergents and glycerol can be used, however, we request that you wash your beads with wash buffer 2X followed by 2-3 washes with wash buffer re-made to not contain detergent or glycerol. Once these additional steps are performed all the liquid can be removed and the beads can either be frozen and shipped to SPARC or proteins can be eluted off.

Gel bands should be shipped at room temperature or at 4^oC.

In-solution or liquid protein samples should be frozen and submitted on dry ice.

Lyophilized proteins and/or peptides can be shipped at room temperature.

Serum/plasma/tissue samples should typically be frozen and shipped on dry ice.

We will provide quotes for individual projects. For quotes for budgeting purposes please contact the Facility Manager at least one week in advance of your deadline. For quotes to generate a PO for samples about to be submitted, submit a request in QReserve and then email the Facility Manager to request a quote.

Collaborators will be invoiced after the project is complete via email, usually at the end of the month during which the work was completed. Payment can be made through internal cost centre for SickKids investigators and by purchase order, wire transfer, or credit card for external investigators.

SPARC operates on a cost recovery basis and is not subsidized by the hospital. Recovered expenses are applied directly to support operation of the facility, including staff salaries, consumables such as analytical columns and solvents, and maintenance services on mass spectrometers and other instruments.

This depends on what technical platform is used for analysis, and taking into account staff time and prioritization of projects. For amino acid analysis by HPLC and proteomics projects, we strive to provide data within 2-3 weeks, but larger projects or unforeseen maintenance issues occasionally necessitate extra time. For small molecule mass spectrometry and immunoassay projects, turnaround time varies depending on current demand and the number of samples. We will do our best to give you an estimate once the samples arrive.

We strive to keep RAW data as long as we can, but encourage users who would like their RAW data to ask for it at the time of experiment.  We will store RAW data for at least one year from the date of your experiment.

Searched data will be kept for six months from the date of your experiment.  We encourage you to treat your data file like any other file, and back it up in multiple places.

SPARC collects fees to offset operating costs. Payment of cost-recover fees is separate from considerations of co-authorship for intellectual contributions to studies that will be published and/or presented.

In cases where intellectual contributions and/or technical assistance are provided by Dr. Moran and SPARC researchers, co-authorship on publications and acknowledgement in presentations will be appropriate, in accordance with accepted scientific practices, journal guidelines, and collegial standards. Investigators are expected to engage Dr. Moran during manuscript preparation to ensure accurate description of methods, interpretation of data, and appropriate attribution of contributions. SPARC does not require the establishment of formal collaboration agreements.