Sensitivity of the HPLC analysis system is the main component in determining the sensitivity of the sequencing system. Preparation of Sample for Sequence Analysis Soluble protein or peptide A soluble sample of peptide or protein can be sequenced directly on the automated protein sequencer. The sample is applied to a polybrene treated glass fiber filter. Polybrene is a polymeric quaternary amine. Golves should be worn, as many contaminants on the skin can be detected in sequence analysis.
Dust is mostly protein, and is also a problem.
All reagents used should be of the highest purity available. All glassware and tubes, etc., should be scrupulously clean. Total volume of the sample to be sequenced should be kept minimal. A good procedure for electroblotting onto PVDF membranes is described in chapter 10 of Current Protocols in Protein Science . As a general rule of thumb, if the stain can be seen, then there is probably enough sample present for sequence analysis.