ABSTRACT
Translation is a central process in gene expression and the development of efficient in vitro translation systems has been the focus of scientific efforts for many decades. The production of translation-competent lysates originating from human cells or tissues remains challenging, mainly due to the variability of cell lysis conditions. Here we present a robust and fast method based on dual centrifugation that allows the preparation of cytoplasmic extracts from human cells that efficiently translate mRNAs in a capped and IRES-mediated way. We optimized lysate preparation and in vitro translation conditions and show that dual centrifugation allows the production of human lysates under detergent-free conditions and is ideally suited to produce ample amounts of human translation-competent lysates.
Competing Interest Statement
The authors have declared no competing interest.