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Center for Protein and Nucleic Acid Research

Protein Sequencing

Protein Sequencing Order Form

Edman Degradation is performed using a ABI Model 494 sequencer. There is a charge of $400 for the first three residues and then $75/residue for subsequent residues. Unfortunately, we cannot offer gel and transfer services at this time. The protocol of running an SDS gel and transferring to PVDF can be obtained here, or you can use the Invitrogen system. We would like to have a minimum of 25 pmol of material for sequencing. For a good signal, typically, 100 pmol of material is required. Typical turnaround for one sequence is about a week.

Tips, Links, & Protocols

We accept protein samples in either of two ways: in solution or blotted on PVDF membrane (PVDF ONLY, nitrocellulose or other membranes will severely damage the instrument). In preparing samples, avoid all amine containing buffers (i.e., Tris, HEPES, Glycine) as these will react with the reagent used in the sequencing reaction and result in no useable data.

For detailed instructions on how to prepare your sample click here.

Download "Preparing Samples for Protein Sequencing, A Newcomer's Guide" for detailed information about preparing samples and the type of instruments we use. (377kb .pdf file)

If you are doing your own transfers download this file for the recommended protocol and PVDF membrane.

In-Gel Digest Protocol

For information regarding the preparation of samples for protein sequencing, please see the Applied Biosystems web page.