Let's say I forgot to add the polymerase enzyme and I only realize this after the run has gone half way, could I stop the reaction to include the pol...would I get amplification if I reprogram afterwards and do a run?
I agree with Utsa; however, you're definitely increasing the chance of seeing nonspecific binding since nothing was amplified after your initial denaturation step. Aside from possibly getting a number of different bands than your desired product if you just continued the cycling after the addition of the polymerase, everything should still work out in theory just not as efficiently.
Ideally, you should restart the program from the beginning, and you should be fine. I hope this helps.
Hi there,
If the polymerase is missing, nothing much happens apart from denaturation and hybridization (maybe some ATP and dNTP hydrolysis). From a synthesis point of view: nothing. So stop the running reaction and then redo from start after polymerase addition. It should work. Not as good as the normal reaction if ATP/dNTP hydrolysis during the unproductive reaction is significative.
Since DNA polymerase is a missing component, the amplification would have not taken place as Max and Utsa mentioned. If programmed is run after addition of polymerase, the multiple bands of different molecular weight will appear along with the band (product) of interest.
If you are trying, i suggest you to upload the AGE pic after running the experiment.
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