I am annealing 37 DNA oligos together but i would like to stop it in certain time.
i tried to add loading dye to stop it but it did not worked.
Do you know how can i stop it?
How are you annealing? Most protocols call for heating the oligos to 94 degrees for 2-5 minutes followed by gradual cooling. The strands actually anneal as the sample cools. why do you need to stop the reaction at a certain time?
i would like to check annealing rate over the time in the specific buffer on ice. I do not anneal them like as routine protocol. if i run the sample in the gel, i will stop it but for 8 samples i cannot run different 8 agarose gel.
Hi Naghmeh,
If I understood your question, annealing means you are using some sort of Enzyme. Right? Just like PCR? In that case, if the heating protocol is not suitable for your cause, I would suggest that you can stop the reaction by poisoning the enzymes. It doesn't always mean like add an inhibitor to the enzyme (IF YOU ARE USING AN ENZYME THOUGH). You can simply drop a PhD changing solution and i sure the enzyme will stop annealing and you can get the "stopped" amplicon! Hope this answer was helpful
Best,
Somnath.
Hi Somnath, No. i am not using any enzyme. i just mix two short oligos in the HEPES buffer and incubate it on ice for different time. now, i want to stop the reaction after 3 min, 6 min, ..... which enzyme can i use for this aim?
Hello, Naghmeh Azadfar, please check linked article. http://www.nature.com/articles/srep05464
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