I was wondering what method should i use if i want to evaluate if a gene is being expressed or not. The first method that came to my mind was Western blot , but i don't know if i could use others. I wanted to know if western blot , specifically , is a good method to evaluate that and , also , if there other methods equally good to evaluate gene expression.

edit : I edit my question and asked about Western Blot , specifically , and asked about more general methods do evaluate gene expression. I don't really know why my question got down voted that much , I apologize for any inconvenient.

  • 2
    $\begingroup$ "Best" depends on what you are doing. For over expression of a heterologous gene in E. coli, I'm content with a dark band of appropriate molecular weight on SDS-PAGE that is not present in the control. That's certainly better than a western in this particular case. $\endgroup$ – canadianer Jul 12 '17 at 19:09
  • $\begingroup$ Perfectly good question! Lots of possible options depending on cell type and whether we are interested in RNA levels or Protein levels. The question might be improved by either asking for a specific system or explicitedly ask for general methods. $\endgroup$ – Jeppe Nielsen Jul 14 '17 at 0:02
  • $\begingroup$ Thanks @JeppeNielsen i edited my question. I also do think this is a perfectly good question. $\endgroup$ – user158657 Jul 14 '17 at 0:33

If you really want to know if your gene is being expressed up to protein, then go with western blot. mRNA based methods are good, but they will be limited in the sense that they will not account for RNAi.

This seems to be a good reference on the subject of RNAi.

| improve this answer | |
  • 2
    $\begingroup$ RNA based methods, while certainly useful in many circumstances, won't account for any kind of non-degradative post-transcriptional regulation. $\endgroup$ – canadianer Jul 12 '17 at 19:14
  • $\begingroup$ Sorry, I should have written RNAi. Take a look here $\endgroup$ – Luís Muniz Jul 12 '17 at 21:01
  • $\begingroup$ Thanks luis. One more thing : If someone says "express" , they mean the production of a certain protein or if the protein is working properly ? I think i just got confused about this concept. $\endgroup$ – user158657 Jul 13 '17 at 2:00
  • 1
    $\begingroup$ @user158657, I found some questions on SE that might be interesting for you. Please check: Q1, Q2, Q3, Q4, Q5 and Q6. I would only add that, personally, I do not think that it is wrong to use gene expression when referring to gene transcription as it is commonly used in the literature. But it is certainly inaccurate. $\endgroup$ – Luís Muniz Jul 13 '17 at 17:20
  • $\begingroup$ @user158657, you could also check these references on non-coding RNAs and epigenetics: ref1, ref2 and ref3. $\endgroup$ – Luís Muniz Jul 13 '17 at 17:20

Not the answer you're looking for? Browse other questions tagged or ask your own question.