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All mixtures were digested using the following protocol.
The trypsinization reaction was stopped with soybean trypsin inhibitor and the mixtures were digested with DNase I.
Mating mixtures were digested with glusulase overnight to eliminate nonspore cells.
The reaction mixtures were digested with proteinase K and then subjected to agarose gel electrophoresis.
A total of 200 μg serum protein mixtures were digested in solution as described above.
To isolate plasmids of the mutants, the PCR mixtures were digested with DpnI at 37°C for 3 h and transformed into electrocompetent XL-10 Gold cells.
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For LC/MS, polypeptide mixtures are digested with trypsin; the peptides are bound to a chromatographic column, eluted with a continuous gradient of acetonitrile and ionized by electrospray directly into either a time of flight or ion-trap mass spectrometer.
For BpmI restriction digestion analysis of DMPK PCR products, 4 μl of PCR mixture was digested overnight with restriction enzyme BpmI (NEB) in a total reaction volume of 20 μl at 37°C.
The trapping reaction mixture was digested with trypsin and samples were analysed by LC-MS-MS.
After separation of the nanoparticle protein complex from plasma, the protein mixture was digested, and peptides were analyzed by nanoliquid chromatography Orbitrap LTQ-XL mass spectrometry.
The mixture is digested on hot plate till it becomes colourless.
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