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The mode of the distributions for presumed tuberculous infection given by the mixture analyses is 15 mm for both datasets.
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Digestion was stopped with 0.25 % TFA and the peptide mixture analysed by MudPIT [ 34].
The mixture was analysed by SDS PAGE and phosphorylated Rpb1 from Pol II was detected by autoradiography.
The mixture was then analysed on a SDS-PAGE gel and the band of the crosslinked complex was excised for trypsin/chymotrypsin digestion followed by mass spectrometric analysis.
After gel filtration on a NAP-25 column (Sephadex G25-DNA grade; Pharmacia) with water as the eluent, the crude mixture was analysed by using a Mono-Q HR 5/5 anion exchange column, after which purification was achieved by using a Mono-Q HR 10/100 GL column (Pharmacia) with the following gradient system: A=10 m m NaClO4 in 15%% CH3CN, pH 7.4, B=600 m m NaClO4 in 15%% CH3CN, pH 7.4.
The reaction proceeded with enough chemoselectivity to identify the desired product from the mixture by the reliable guidance of spectroscopic analyses.
By this time the mixture will thicken.
10 μl of the amplification mixture were analysed by electrophoresis in 1.5 agarose gel and visualized by UV light after ethidium bromide staining.
The product mixture was analysed by chromatographic and spectroscopic methods.
Twenty per cent of the peptide mixture was analysed by nano-liquid chromatography tandem mass spectrometry using an LTQ Velos-Orbitrap MS (Thermo Scoupledic) coupled with an Ultimate RSLCnano-LC system (Dionex).
The peptide mixture was analysed by LC-MS (Agilent 6300 Series Ion Trap LC, Mass Systems).
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