Click here to go to the tutorial
Unsurprisingly, my most common approach to proteome abundance measurements by mass spectrometry is data independent acquisition (DIA). Specifically I’ve been using the chromatogram library approach (Searle et al 2018) because, compared to spectral library-based approaches, it doesn’t take a lot of extra work. I just prepare my samples as usual, then pool a few uL of each sample into a “library” or consensus sample. I queue up my single-shot experimental samples, then I acquire the pooled library sample with multiple injections, each time spanning a 100 m/z range (gas phase fractionation, GPF) with very narrow isolation windows.
The next step up is to search the narrow window, GPF multi-injections against a spectral library. Recently, a team of researchers released “Prosit”, a tool to predict spectral libraries. Using Prosit predicted spectral libraries to search GPF chromatogram libraries gives detection numbers a boost (Searle et al 2019). Because it’s so easy to use predicted spectral libraries, I’ve been doing it for all my projects.
The tutorial above is a work in progress, so let me know if you have questions or suggestions to improve it!
Learning to code, coming from an experimental background, can be a frustrating and intimidating experience. Although there are many free courses online that will teach you the basics of Python and programming, the best way to improve is to simply practice. The best practice is a project you are personally motivated and passionate about. For graduate students, this might be a component of your thesis or a side project that complements your research interests. Coming up with an interesting project might be daunting at first, however, so here are some resources for quick, achievable practice problems to help keep you coding.
Practice mathematical/computer programming problems
Python basics with Rosalind.info
Beginner bioinformatics with Rosalind.info
Check your code style online
Students, teachers, parents — welcome! Thank you for your interest in following up on the Expanding Your Horizon’s workshop! Here’s a summary of what we talked about today, along with many links to videos and sites where you can learn more about gene editing.
For the 2017 Expanding Your Horizons event in Edmonds, WA, the University of Washington’s Women in Genome Sciences group created a workshop around the topic of gene (genome) editing (find the lesson plan here, and the presentation here). Our goals for this workshop were to communicate the key terms and concepts of gene editing, and apply a scientific thought process to some familiar and not-so-familiar problems. Read more about our day below.
Continue reading “Digging Deeper into Gene Editing at Expanding Your Horizons (Edmonds, WA)”