u/Best-yogurt7481

I have been struggling for too long with my job in research, and I am now realising that the impact this has on my mental health is too strong. I love science but maybe I'm not done for it. It's hard to face this disappointment everyday-.

I think I need psychological support but I would like to speak to someone that understands academic dynamics. I could not find anything online. Do you know if German universities o privates offer something of that sort?

Where can I find some help?

I moved here a month ago. I haven't had much time to explore the city due to crazy working hours, no car and not speaking even the most basic german.

Bit I am enjoying a lot the river and the gardens, and I have the good impression af a livable and quiet city. So far it's been really nice here. I would also say that the spring weather helps.

I am curious to know what you like (or not like) of it? From every aspect: cultural, architectural, social, etc.

Hi. I'm quite new to the protein purification world. I am having problems in expressing a protein. Would there be someone able to tell me what I did wrong?

- I transformed my DNA in BL21(DE3) rosetta strain and directly inoculated the 1mL post-transformation into 100 mL overnight culture with antibiotic - skipped the plating step.

- this morning the culture was turbid. I pelleted as recommended from the protocol I'm following (I would say there was a more than decent pellet).

- here comes the issue: I resuspended into 4 mL and inoculated in 4 L media with antibiotic, etc. After 4 hour OD600 is 0.02. So nothing. Did I create a massive bottleneck, should I have resuspended the pellet in more volume? I did it once already in the past and it worked. I only hope I did not waste 4L media and created another delay in this process. Is there be any chance for the grow to start at some point?

Thanks to everyone who can give me some advice!