1) 32 percent on an exam can be a great score
2) 4 mountain dews do not equal a full night of sleep
3) The phrase “if you put it on a slide, you have to know it.”
4) Learn a whole lot about very little
5) Personal hygiene is not universal
6) Snacks offered before a guest lecture do not count as a meal
7) Don’t end a first date with ‘I love you.’
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What lessons did you or are you learning?
Protein crystallization is influenced by a number of variables. Initially, one must decide what type of setup will be used when attempting to grow a protein crystal.
A common method is hanging drop, which utilizes a process called vapor diffusion. The benefit of vapor diffusion is that it allows for the increase of protein and precipitant concentrations. The concentration is increased due to vapor leaving hanging drop to reach equilibrium with its surroundings.
Hanging Drop Setup:
Note: This setup does not allow for evaporation to take place although it may be used to encourage crystal growth. The cover slip is often sealed in place using vacuum grease.
The contents of the hanging drop includes your protein with its buffer, well solution and sometimes water. A common setup is to use 2 µl of protein solution + 2 µl of well solution, suspended over 998 µl of well solution (the 2 µl are added from the well solution to the hanging drop).
1) Bio-Rad thermal cycler: The PCR song
2) Eppendorf pipetting: Boy Band
Enjoy the weekend!
Gordon points us to the CCP4 Study Weekend which is going to have a live feed this year– awesome! The show gets on the road January 8th at 9 am GMT (4 am EST). The CCP4 study weekend is taking place in Nottingham, UK this year and as usual has a number of excellent speakers.
The videos look like they will be archived for those who are unable to watch the events live. For those that tweet @pssalgado is going to be using #ccp4 for updates.
If you are struggling with refolding your protein then you may want to take a look at REFOLD. The Refold database currently contains 759 protein entries.
The help pages are really nice in that they explain how to use the database and contain background information. For example, here is a pdf of ‘A practical guide to protein expression and refolding from inclusion bodies.’ Based on the current entries the most common method of refolding is by dilution.
Here is a shot of a couple of search options (note: all the panels are not open)
Graphs are updated nightly to reflect the current contents of the database:
A neat feature of this database is that it allows users to create their own free account which will track their past searches.
The CCP4bb is the most popular electronic mailing list that is related to macromolecular crystallography. The idea would go even further and bring together all bulletin boards (such as Phenix, Coot, PyMol) into one location. This would benefit developers and users in that they would only need to check one location to ask and answer questions.
It is time to move on when:
1) Members have a drawn out (here, here, here, here, here, here, here, here, here, here, here) discussion on attachment size and html formatting in emails which is fine.
Does everyone need to receive each of these emails? No.
Both problems would be fixed by using a forum instead of an email based system.
2) Members are being used as targets for scams (yeah, I don’t know who falls for these either).
3) Have your work email address spammed due to being on the bulletin board.
4) You would not have to depend on others to summarize answers to the question they asked. This would not only save time of those asking questions, but allow others to have access to all the answers.
5) A forum could be organized by topic instead of the entire list receiving every email.
Members of the email bulletin board have already expressed their dissatisfaction in receiving every email.
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What would the topics look like for this new forum?
Protein Cloning, Expression and Purification
Protein Crystallization
Data Collection
Data Processing
Data Refinement
Program Installation Problems
Program Execution Problems
Employment
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The topics will need to be adjusted. It won’t be perfect at first, but will be much, much better.
It is time to make an old system fit the next generation.
Please drop me a line in the comments on what you think about this and if you would join.
As Peter Müller may say Artem is looking for a Jedi Knight.
Protein Science Intern (St. Louis, MO)-000W8
Description from the Monsanto site:
We currently have an opening for a Summer Intern in Structural Biology and protein biochemistry. This is an excellent opportunity for a fledgling scientist to work on challenging projects and to learn advanced skills in protein expression, purification, and crystallization. Working under the guidance of a research scientist the prospective candidate will be given an opportunity to contribute his/her work towards an important project related to Monsanto’s agricultural technologies. The opportunity to publish results in a peer-reviewed journal may also become available. The duration of the appointment is structured to last 8-10 weeks with 40 hr per week commitment during the Summer months of 2010. This position will be located at Monsanto Chesterfield facility in St. Louis.
More details as well as where you need to apply are found on the Monsanto site (for easy searching use 000W8 as the Requisition Number)