Hi guys!
For the lab exercise we designed molecular inversion probes. The first batch we designed didn't work, so we ordered a probe from the original paper and it still didn't work...
The main mechanism is the probe is circularized if it fits to the template.
The single stranded probe is 115 bases long. How can we check if circularization worked without radioactive labeling or expensive stuff?
I was thinking of running a gel - circular 115 bases should run slower than linear 115 bases. Though the concentration is very very low - attomols - so no chance to see on a gel.
Also, just using one primer and making multiple tandem copies (to get 230, 345, 460, ... bases) and subsequent amplification won't work (Polymerase won't kick away the primer)...
Does anyone know an easy way?
-- For the lab exercise we designed molecular inversion probes. The first batch we designed didn't work, so we ordered a probe from the original paper and it still didn't work...
The main mechanism is the probe is circularized if it fits to the template.
The single stranded probe is 115 bases long. How can we check if circularization worked without radioactive labeling or expensive stuff?
I was thinking of running a gel - circular 115 bases should run slower than linear 115 bases. Though the concentration is very very low - attomols - so no chance to see on a gel.
Also, just using one primer and making multiple tandem copies (to get 230, 345, 460, ... bases) and subsequent amplification won't work (Polymerase won't kick away the primer)...
Does anyone know an easy way?
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