diff --git a/assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF analysis of Si EPS and CW.txt b/assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF.md similarity index 91% rename from assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF analysis of Si EPS and CW.txt rename to assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF.md index aecab76d1f9c6b59d5463ac0ab26ccfb520cc54e..11cb825add81b54dcc6900be290467155a9d30ce 100644 --- a/assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF analysis of Si EPS and CW.txt +++ b/assays/MALDI-TOF analysis of Si EPS and CW/protocols/MALDI-TOF.md @@ -1,7 +1,7 @@ -# Analysis of oligosaccharides released from the EPS matrix and CW of S. indica +## Analysis of oligosaccharides released from the EPS matrix and CW of S. indica Freeze-dried EPS matrix (1 mg) or CW preparation (1 mg)isolated fromS. indicawere suspended in 400mLofa2-mM sodium acetate buffer, pH 5.0 (TLE), 2-mM MES buffer, pH5.0 (H. pomatiab-1,3-glucanase), or 25-mM sodium acetatebuffer, pH 5.0 (HvBGLUII) and incubated at 70C overnight.The suspended material was treated with 2.5mLofTLE,2.5mLofH. pomatiab-1,3-glucanase, or 1mLofHvBGLUII inthe respective buffers, as described before, in a total reactionvolume of 50mL. The digestion was performed at 40Cwithshaking at 500 rpm for 16 h. The digestion reaction wasstopped at 95C for 10 min and centrifuged at 11,000gfor 5min at room temperature. -# Mass spectrometrical analysis +## Mass spectrometrical analysis The oligosaccharides present in the prepared samples wereanalyzed by Oligosaccharide Mass Profiling as described(Gu ̈nl et al., 2011). Briefly, the samples were spotted onto adried spot of dihydroxy benzoic acid matrix (10 mgmL–1)and analyzed by MALDI-TOF mass spectrometry (BrukerrapifleX instrument). The machine was set to linear, positivereflectron mode with an accelerating voltage of 20,000 V.The spectra from the samples were analyzed using flexanaly-sis software 4.0 (Bruker Daltonics).