Precipitated P. falciparum protein preparations were dissolved in digestion buffer, digested with trypsin and LysC, and analyzed by LC/LC/MS/MS according to published protocols [22], [23].
The purified samples were dissolved in 100 μl of digestion buffer, and digested at 50°C with 50 U of SfiI overnight.
DNA was isolated (Promega Wizard DNA isolation kit) from tail snips digested in proteinase K digestion buffer (20 mg/ml).
Similarly, tissues (mainly muscle) were digested with Proteinase K and digestion buffer, whereas saliva swabs were spun for DNA extraction from the resulting pellets.
Cells were digested with Zymolyase in the digestion buffer (50 m m Tris-HCl, pH 7.5; 10 m m MgCl2; 1 m Sorbitol).
Briefly, the samples were digested overnight using Proteinase K and a digestion buffer.
After incubating for 15 min, 90 μl of digestion buffer was added and the samples were left to digest overnight at 37°C.
Proteins were redissolved in trypsin digestion buffer, quantified using the BCA protein assay (Pierce, Rockford, IL, USA), and digested with trypsin as described above for the exudate samples.
Eske Willerslev and Philip Francis Thomsen of the University of Copenhagen and colleagues used a solution called a digestion buffer, the recipe for which had been previously developed.
It's thought that the digestion buffer gets inside the exoskeletons through the mouth, respiratory holes and other anatomical features, and through holes made when the specimens were pinned for display.
The denaturing solution was replaced with a digestion buffer using a custom-designed 96-well size-exclusion plate for desalting.
