Nephrite

Use geNorm or NormFinder to find most stable genes ( I would, but geNorm is currently not working in my 64bit Office)
But from my point of view you don't have much to choose from. 18S has the biggest problem of being too abundant, so maybe limiting the primers would help a bit, and then you only have GAPDH and actin, which is know to be problematic, you have even twice.

Ct variation between replicates is not telling you anything about the housekeeping gene's stability (if I understand correctly that those are technical replicates, not biological).

Not really Andrea, I did my MSc in UK, but I couldn't study PhD since it was really expensive and they didn't give full scholarship to non-EU students. The only scholarship available to non-EU students was called ORS or something, but that only paid GBP 7K per year and it was almost half of the GBP 11k university fee. I worked part time at Cafe Rouge in Colchester during my MSc, and at night I guarded a residential house for the elderly. I didn't have to, but I did it because I wanted to be independent and didn't want to receive support from my parents. The pay was not bad to maintain myself, but then I thought I couldn't do the same part time jobs in my PhD. Since I graduated with distinction my supervisor introduced me to one of her collaborators in Giessen, Germany. That time German universities were free (not sure about now). So I came down to Giessen in early 2006. But right after my arrival the supervisor claimed his grant application was turned down and he couldn't support me in the first year of my PhD. I was like what the hell, the guy has articles in Nature and Science, how unlucky I could be. I calculated how much money I needed to survive the first year of my PhD with no funding and it was almost Eur 8K. I was right in the middle of the calculation that I got a full scholarship offer from Malaysia. I immediately packed and jumped into the plane and here I am now...things were pretty good here, they gave me a 3-bedroom apartment, and after 3 months I bought a second-hand car with my savings. The research is not first-class here, but they have this program that they are absorbing forigners to develop their research...Now I am doing my postdoc, the pay is much better than UK's. If I were in UK I would have only got GBP 2500-3000 per month, and that is not enough at all.

Overall I am happy here, but the problem is that the research in here or in my own country won't satisfy me enough. I know I can be better than this, I want to grow and learn more. I know I have the talent to publish better articles. I'm still young and haven't reached the point that I need to sit down and relax...not yet...but it is like the moment people read my CV, and see where I got my PhD from, they turn down my postdoc application...I think they don't even reach the last page where the names of my American referees are....now I keep thinking maybe I should have borrowed money from someone and paid my living expenses in Germany to get a world-class degree....I really regret it....but you never know what's gonna happen tomorrow.

Nephrite, on 11 September 2012 - 06:17 AM, said:

Hi Nephrite,

I look at this differently...you shld be extremely proud of what you've accomplished all by yourself (of course with the help of your professor), despite all the challenges you encountered. And now that you've returned to your country, you should be able to find inspiration in the people that you care about, in the places that you love, in the things that you missed when you were out so this should actually motivate you more. You shldn't cop out otherwise, everything that you've endured would be for nothing. You've got a PhD you worked so hard to get. You lived in a foreign country and survived (how many in your country had this opportunity?)

You're feeling down and very tired now so you're allowed to take a well-deserved rest but once you have regrouped you'd realise that you will never allow yourself to become a useless member of society. Hopefully these feelings of depression and lack of motivation are only temporary but if they persist perhaps you shld also consider consulting a mental health professional.

If you really don't want to do research anymore, as the others have suggested, there are other career directions that you can take. But is the level of research in your home country comparable to the one you left behind? Perhaps there would only be simpler hypotheses to prove, cheaper techniques to employ etc. And are there many PhD degree holders as yourself? If not,then you definitely have an advantage so don't sell yourself short (and cleaning houses shldn't be an option cos you would probably suck at it ). Be strong and good luck.

I applied for 3 academic jobs in my country just yesterday. Wish me luck....Most probably it will be turned down, but let's just hope they approve me....

To add to Casandra's comments above: kudos for doing most of the stuff on your own. On top you have several publications. Double-kudos. This means you are good. And you became even better because you survived. I know of others in your situation who didn't survive and they were lost. In my opinion, you should take a long walk and relax and think what you really-really want to do. Not what society expects you to do. what your parents/partner expect you to do. what do you want to do? I believe that you would make it in research, if you decide that actually, the reason you started a PhD in the first place was the love for research. But, if you want to have money or a family with whom you have a normal relationship based on seeing them for dinner every night, then go to industry/consulting etc. etc. Teaching... hmm does not bring you money, not research satisfaction...maybe the warm feeling inside from molding the future generation, really, most of them don't want to be molded anyhow But this is just me. In the end of the day, a PhD is a confirmation of the mind/spirit (smth they keep on telling us over and over in my university city in all the speeches) My bottom line is: a PhD title is like a title that you are an extremely educated/intelligent person. You can do whatever you want, from writing articles in the science section of your daily newspaper or consulting for patent lawyers or human resources in a company that has an R&D department based on life sciences/chemistry. You are basically an expert on nothing and everything.

With going back to your own country, reply at Casandra: many countries, like China, might have these programs to support the people to come back home and do research, but as far as I have heard for both China and my home country, Romania, they are a joke. They just sound good on paper but they crumble after the first wind of reality. I know of people going back to China for a professorship and doing only lecturing and not having a proper lab until they gave up and gone back to UK for a postdoc. aka from professor to postdoc. And about Romania, among my network of friends that studied abroad and consider going back, the rumor has it that it is still not the time: many have gotten tricked in going back and not they regret it. So: it might sound nice in principle, but in practice, it does not work. And anyhow, I am now in Germany for long enough time to actually apply for the citizenship and stay here forever and ever. And I will just do so, because as you have said: they make it easy for us to get work permits and residence permits. I got both of them with no problem: fill in a form and both of them are without end.

casandra, on 11 September 2012 - 07:50 PM, said:

Casandra, you made my day

Nephrite I understand you totally. I am finishing the first year of my postdoc in a couple of months, and after that I don't know what's gonna happen. I feel lost. I work in a foreign country too, and I can't go back to my own country either because I have been away for nearly 10 years and I don't know anything about the industry or acadmic culture in there. I am not attached to any organization and if I go back I will have to be unemployed for some time until I evaluate my foreign degrees and find a junior lecturer position. It's a shame really.

I think everybody wants to be useful in the society somehow. In our field of science maybe being useful means publish article in high impact factor journals and get a lot of citation. But I didn't have any article until few years ago, so I kept asking myself what if I get hit by a car tomorrow and my life ends with no outcome? I told myself if I don't have articles, then maybe there is another way to be useful...and I found it....I joined Bioforum...first I joined to find my own answers, but later I started answering other people's scientific questions. I started with small questions, questions that I knew the answers for, and left the difficult ones to the moderators or veterans. Towards the end of my PhD I became more and more addicted to this, and I could answer more questions related to the methods and protocols that I use in the lab everyday. I have been here for many years now. If not every day, I have visited this forum every week. I joined when there was no 'like' button , and now I am a moedrator myself. I do this voluntarily. We don't earn anything, but it satisfies me enough. My wife used to get mad at me why I spend too much time here, for something that I don't get salary for, but now she understand my passion and doesn't argue anymore. Who knows, maybe one of the people who I helped becomes the greatest scientist of all time. Maybe he won't remember me, or I won't remember him, but I helped him in his way to success. and that counts.

I don't want to sound philosophical, but I always remember what Isaac Newton said that 'If I can see further, it's because I am standing on the shoulder of giants'. He was right, and everything counts. If there is a God (that I am not sure) he sees everything, and the universe will take record of what we do....That's how I see it.

what about other aspects of a biotech/pharma company? Sale, patents, technical support, consultancy, recruitment...?
Companies often search for PhD educated people for these positions.
I met several people who after a PhD or a post-doc left academia and research for a different job in the life science field.

you can try for another post-doc position elsewhere or join an industry as a post doc

Template-dependent primer-dimers

I just had an idea, maybe you already thought about this or even wrote about it somewhere in the forum, but this striked me just now, so I'm writing it here because it's a sticky topic and someone may be interested. (or maybe as said in czech, I just discovered America - something already well known).  

While using SYBR method (mainly UPL-designed primers that eventualy were used without the probe) I quite often encountered template-dependent primer-dimers, NTC was positive for a product with lower melting temperature, probably dimers, but there were no dimers in any sample, just the specific peak. Since it wasn't interacting with sample measurements I just let it go, but I always wondered, why there are dimers in NTC but not in samples, when there all the same except template?

Just now i realised what actually is different in NTC, the amount of free Mg+ ions!
DNA captures Mg+ ions (thanks to someone who mentioned it somewhere) and decreases the free pool of them. But in NTC there is no DNA and there are more Mg+ that makes the mix less stringent. In less stringent enviroment, the dimers form but they don't in samples with a higher stringency.
This could be the perfect reason why there are template-dependent dimers. What do you think, makes sense?