Author: Conceição-Neto, Nádia; Theuns, Sebastiaan; Cui, Tingting; Zeller, Mark; Yinda, Claude Kwe; Christiaens, Isaura; Heylen, Elisabeth; Van Ranst, Marc; Carpentier, Sebastien; Nauwynck, Hans J.; Matthijnssens, Jelle
Title: Identification of an enterovirus recombinant with a torovirus-like gene insertion during a diarrhea outbreak in fattening pigs Document date: 2017_9_8
ID: kgoczioe_21
Snippet: DTT was added to 20 lg of protein extract up to 20 mM of final concentration and incubated for 15 min. Then, iodoacetamide was added to the mixture up to a final concentration of 50 mM and incubated for 30 min in the dark. Then the sample was diluted 3 times in 150 mM ammonium bicarbonate. For protein digestion, 0.2 lg/ll trypsin was added and incubated overnight at 37 C. Samples were acidified with trifluoroacetic acid (0.1% final concentration).....
Document: DTT was added to 20 lg of protein extract up to 20 mM of final concentration and incubated for 15 min. Then, iodoacetamide was added to the mixture up to a final concentration of 50 mM and incubated for 30 min in the dark. Then the sample was diluted 3 times in 150 mM ammonium bicarbonate. For protein digestion, 0.2 lg/ll trypsin was added and incubated overnight at 37 C. Samples were acidified with trifluoroacetic acid (0.1% final concentration) and purified with Pierce C18 spin columns (Thermo Scientific) according to the manufacturers' instructions. Peptides were eluted with 40 ll and then evaporated using a speedvac.
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