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After washing of the trapping column with 0.1% trifluoroacetic acid (TFA), the peptides were eluted (flow rate300 nl/min) from the trapping column onto an analytical column (Aurora C18, 75 m ID, 250 mm long, 1.7 m particles, heated to 50 C, Ion Opticks) by 60 min linear gradient program (3-42 % of mobile phase B

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