Solutions
Design Solutions
Gene Construction
Gene Library Filters
Optimization and Analysis
Applications
Gene Library Filters
CODA utilizes two types of Library Filters:
  • Quality Control Filter
  • Solubility Filter

Library Quality Control

Gene variant libraries typically contain a high percentage of insertions or deletions leading to out-of-frame sequences.. CODA’s unique quality control system, pCODA-IN, provides a biological filter that can provide libraries with 60- 90% in frame sequences.

Selection principle:

  • pCODA-IN in-frame library selection
  • selects regardless of folding, solubility of function of the translated fusions
  • Sec fused to selective marker
  • Self-catalyzed excision ensured or antibiotic resistance conferred if the insert remains in frame

 

 
   
Solubility Selection Filter – SolSelect

CODA has developed a high-throughput solubility screening and selection system to identify and clone soluble variants from rationally designed libraries of any protein of interest in E. coli. Taking advantage of CODA Genomics’ thermodynamic Hot Rod™ gene assembly technology, gene deletions are then constructed via multiplex PCR producing deletion libraries with 95% coverage of the possible constructs.

   

CODA constructed a SolSelect combinatorial library (CL), with minimal number of concurrent amino acid changes, for the SAM domain (87 amino acids) of TEL (Translocation Ets Leukemia) by thermodynamic assembly of specified molar ratios of wild-type and mutant oligonucleotides and subsequent fragment library assembly. SolSelect CL TEL-SAM was sequentially qualified by pCODA-IN in-frame selection.  It was then  inserted into pCODA-OUT and used to transform E. coli TOP10 for screening and selection of soluble clones.