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17 June 2013
From 2013.igem.org
Today, we assisted Madina (one of our advisors) to do the SELEX cycle 8 (selecting aptamers against CEA) and to finish cycle 3 of SELEX (selecting aptamers against ECPKA). For SELEX cycle 3 we have finished with precipitation of DNA aptamers and have put them store at -20degC, to use it in SELEX 4. For SELEX 8 we did the following steps:
- Denaturation of dsDNA from SELEX 7
- Pre-wetted (passing DNAs through pre-wetted nitrocellulose membrane)
- Binding (incubating denatured DNA with the target protein CEA)
- Washing (passing binding reaction mixture through the membrane using washing buffer)
- Elution is done to degrade protein, and to remove it from DNA
- Precipitation (eluted DNA was diluted with water, and then 100% ethanol, ammonium acetate and glycogen were added). The mixture was incubated at -80degC for one hour. After that, precipitation was done.
- PCR to make dsDNA
- We checked PCR product with 6% TBE gel.
