is there is a method to test RNA dependant RNA polymerase without using a radioactive material?

Question

RNA dependant RNA polymerase use the RNA as a tempelate to senthesise a new strand . But the problem in my work is that i do not want to use any radioactive material to asses the activity of this enzyme . SO can any one tell me how to do that with involving a reference
thank you very much for your help..
my email adress is : [e-mail]

Answer 1

I don't have a specific reference for you, but your research adviser should be able to point you in the right direction. Based on your rather general need to 'assess the activity', I can make two suggestions you look into:

When assessing enzyme activity, one often uses quantitation of either product formation or substrate elimination. You can probably use UV spectroscopy to quantify RNA strands vs. RNA bases.

If you want to label your enzyme, why not use a fluorescent molecule? All you will need then is a fluorescent microscope. This is a method used in many cell biology labs, like labeling actin molecules to observe sppindle fiber movement during mitosis or labeling the DNA in an egg and a sperm cell and then observing the combination of these DNA upon fertilization.

To find a reference, why not go to google images and type in fluorescent cell or something like that, and then go to the URL that has posted their images.

For the UV spec protocols, I would look at molecular biology lab manuals.

Answer 2

The answer is A - it's part of the process of transcription. (B is translation, C is cell death, D is an artificial laboratory method of replicating DNA. None of these need to make an RNA polymer therefore no RNA polymerase.)

Answer 3

I have not heard of this polymerase before. However, I guess you can design a Polymerase Chain Reaction to try it out.