In Vitro Directed Evolution of Enzymes Expressed by E. coli in Microtiter Plates
A method is described for using 96-well plates to prepare libraries of Escherichia coli cultures for screening a library of gene variants. This approach bypasses colony-picking to allow standard molecular biology laboratories to carry out directed evolution efficiently with a 96-well plate-reader and multichannel pipettes. Initial screens are applied to cultures that are rapidly prepared by diluting transformed cells so that an average of four cells starts each culture. Subsequent screens are used to isolate individual enzyme-expressing clones that exhibit activity higher than the parental clone. The outlined method also includes guidelines for preparing a library of gene variants and for optimizing a screening method.
- 石蜡加填充剂 Paraffin Embedding
- Isotype-specific immunostaining with mouse primary antibodies on mouse tissue
- 犬D-乳酸(D-Lactate)酶联免疫分析
- 南味花生糖
- A Modified Coupled Enzyme Method for O-linked GlcNAc Transferase Activity Assay
- 复制(replication)
- DNA电镜技术及其应用
- Purification, Reconstitution on Lipid Vesicles, and Assays of PDE6 and Its Activator G Protein, Transducin
- 沙眼衣原体诊断研究进展
- 儿茶酚胺类代谢