Kia ora, I am designing a SELEX protocol to purify a oligonucleotide sequences towards a protein conjugated to magnetic protein G beads. I want to know whether heat elution of specific sequences from the target or chemical elution i.e. using an elution buffer or NaOH, EDTA etc. would be better. I have seen both used in the literature but wonder if anyone would be able to provide any recommendations as well as any papers they found informative.
Thanks for your help,
Rebecca
The conventional method has been heat-based elution. The buffered elution is better. There are other factors to consider, temperature, concentration, media, denaturation extent, heat-lability, length and nature of the sequence, etc.
Check these: Article Small-Molecule Binding Aptamers: Selection Strategies, Chara...
https://application.wiley-vch.de/books/sample/3527342672_c01.pdf
https://www.researchgate.net/topic/SELEX
For me elution is better but also you need to heat to eluate your sequences
For me I had to change the method so as not to immobilize the target because when heating to 90 C my target degraded and the sequences obtained were specific to the degradation product not to my initial target. so I opted for the selex without immobilization and extract my sequences by PCI
But if heat does not work? Not give any band of eluted Aptamers on Pilyacrylamide gel. Then what would be the other better plan to elute the bound aptamets with imbolilized protein targets??
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