What are the molecular techniques that I should use to evaluate the replication, transcription and translation in vitro?
http://ecotoxicologia-bioensayos.blogspot.cl/2015/12/biothecnology-practice-university-of.html
For other readers: it looks like he is doing bacterial transformation with the pGLO vector with GFP.
I don't know about replication but transcription: RT-qPCR and translation: Western blotting.
The answer will be a manual of some hundred pages. Better ask more specific questions.
Since these methods are easily covered everywhere online and in textbooks, I will assume you are looking for updated methods and techniques. So with that...
Transcription rates cannot be measured by normal RT-qPCR through total RNA extraction, that will only give you steady state levels relative to some arbitrary baseline.
If you actually want to measure transcription rates perform a nuclear run on as would be done in GRO/PRO-Seq, and then do qRT-PCR or sequencing.
http://www.sciencemag.org/content/322/5909/1845.long
¿ If someone could help me resolve this box located in the Word document someone could help me resolve this box located in the Word document?
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