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Center for Protein and Nucleic Acid Research

Protein Sequencing Costs and Instructions

Rates are based upon the number of amino acid residues sequenced. A standard initial charge of $200 for the first 3 amino acid residues and $35 for every additional amino acid residue.

Protocols

Typically, 25 pmol of protein are required for a good signal.

Protein samples may be submitted in one of two ways. In solution or bound to a PVDF membrane.

1. In solution, the sample should be in a volume of about 30 uL. The sample will be bound to a PVDF membrane, and the membrane placed on the instrument for sequencing. An additional charge of $25 will be applied for this service.

2. The second method, the sample can be electroblotted to a PVDF membrane from an SDS gel. The band visualized by coomassie staining, excised, and submitted in a tube. The excised protein band on the PVDF membrane will be loaded onto the instrument for sequencing. The band should be no bigger than about 7 mm x 7 mm. The coomassie staining will not affect the sequencing. Keep in mind when deciding how much sample to load on the SDS gel that the efficiency of electroblotting can be anywhere from 40-90%, so if you need 25 pmol of sample to be on the membrane you should load roughly around 50 pmol of sample on the gel. YOUR SAMPLE MUST BE ON PVDF MEMBRANE. If you use some other type of membrane (like nitrocellulose), it will damage the instrument badly. This will require a service call on the instrument to replace fluid lines, glass cartridges, and the HPLC column. The costs could be as high as $2,000. The facility will have no choice, but to pass these costs along to your lab, so please make sure your sample is on PVDF.

For peptides, they should be submitted in solution. They will be bound to microTFA filters and placed on the instrument for sequencing. An additional charge of $25 will apply.

If you have any questions feel free to call 4-9941 or email peptide@scripps.edu.