High-Throughput Purification of Combinatorial Libraries II:  Automated Separation of Single Diastereomers from a 4-Amido-pyrrolidone Library Containing Intentional Diastereomer Pairs

2004 ◽  
Vol 6 (4) ◽  
pp. 478-486 ◽  
Author(s):  
Mark Irving ◽  
Clinton A. Krueger ◽  
Janice V. Wade ◽  
John C. Hodges ◽  
Kyle Leopold ◽  
...  
2020 ◽  
Vol 21 (22) ◽  
pp. 8774
Author(s):  
Natalia Komarova ◽  
Daria Barkova ◽  
Alexander Kuznetsov

Aptamers are nucleic acid ligands that bind specifically to a target of interest. Aptamers have gained in popularity due to their high potential for different applications in analysis, diagnostics, and therapeutics. The procedure called systematic evolution of ligands by exponential enrichment (SELEX) is used for aptamer isolation from large nucleic acid combinatorial libraries. The huge number of unique sequences implemented in the in vitro evolution in the SELEX process imposes the necessity of performing extensive sequencing of the selected nucleic acid pools. High-throughput sequencing (HTS) meets this demand of SELEX. Analysis of the data obtained from sequencing of the libraries produced during and after aptamer isolation provides an informative basis for precise aptamer identification and for examining the structure and function of nucleic acid ligands. This review discusses the technical aspects and the potential of the integration of HTS with SELEX.


2007 ◽  
Vol 74 (4) ◽  
pp. 1190-1197 ◽  
Author(s):  
Christine Nicole S. Santos ◽  
Gregory Stephanopoulos

ABSTRACT We present the development of a simple, high-throughput screen for identifying bacterial strains capable of l-tyrosine production. Through the introduction of a heterologous gene encoding a tyrosinase, we were able to link l-tyrosine production in Escherichia coli with the synthesis of the black and diffusible pigment melanin. Although melanin was initially produced only at low levels in morpholinepropanesulfonic acid (MOPS) minimal medium, phosphate supplementation was found to be sufficient for increasing both the rates of synthesis and the final titers of melanin. Furthermore, a strong linear correlation between extracellular l-tyrosine content and melanin formation was observed by use of this new medium formulation. A selection strategy that utilizes these findings has been developed and has been shown to be effective in screening large combinatorial libraries in the search for l-tyrosine-overproducing strains.


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