Abstract:
:Ribosome profiling is a powerful tool for characterizing in vivo protein translation at the genome scale, with multiple applications ranging from detailed molecular mechanisms to systems-level predictive modeling. Though highly effective, this intricate technique has yet to become widely used in the microbial research community. Here we present a streamlined ribosome profiling protocol with reduced barriers to entry for microbial characterization studies. Our approach provides simplified alternatives during harvest, lysis, and recovery of monosomes and also eliminates several time-consuming steps, in particular size-selection steps during library construction. Furthermore, the abundance of rRNAs and tRNAs in the final library is drastically reduced. Our streamlined workflow enables greater throughput, cuts the time from harvest to the final library in half (down to 3-4 days), and generates a high fraction of informative reads, all while retaining the high quality standards of the existing protocol.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Latif H,Szubin R,Tan J,Brunk E,Lechner A,Zengler K,Palsson BOdoi
10.2144/000114302subject
Has Abstractpub_date
2015-06-01 00:00:00pages
329-32issue
6eissn
0736-6205issn
1940-9818pii
000114302journal_volume
58pub_type
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