Example Isolation of Moenomycin A-specific Aptamers

In this section, we utilize the selection of moenomycin A-specific 2 -modified RNA aptamers by affinity chromatography [28] to give a more detailed overview of a SET EX experiment. 

The released 2-thiopyridone was quantified by UV spectroscopy (343 rnn) to determine the amount of immobilized moenomycin A. 

The moenomycin-sepharose was stored in selection buffer at 4 °C and could be used for about 2 weeks. 

The reaction was carried out in five 100 pL mixtures containing 1 x PCR buffer, 200 pM of each clNTP, 1 pM primer A, 1 pM primer B, 2U DNA polymerase, and 10 pL of the RT mixture. In the first SET EX round, the ssDNA pool was used as the template. 

The dsDNA product was ethanol-precipitated and dissolved in ddH20. 

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