Hello all!
I am currently aiming to extract RNA from human macrophages infected with M. tuberculosis and facing some issues related to RNA extraction and cDNA quality.
To summarize.. Is it possible/recommended to use in the same reaction both Random hexamers and oligo(dT) in an attempt to improve the results of the method?
To clarify, I am aiming to evaluate both RNA from the macrophage and the bacteria.
Thank you for your attention!
Hello @Rachel and @Shambhu!
Thank you very much for your advice and attention!
I will make sure to try it out in my next RT.
Yes, you could add both things in one reaction mixture but as described above for random primers you should incubate at room temperature and for oligo dT its mostly 42°C -45°C. You could perform both steps serially and go ahead with next step. It works better for me always.
I think you must choose the suitable primer oligo dT or random primer
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