Batch barcode generator7/26/2023 ![]() 10 conjecture that the cost of gene synthesis could become on par with oligo pools (1 USD per 10 3 to 10 5 bp), which would allow for DNA barcodes to be used in practical applications in which a designed DNA sequence and unique DNA barcode are designed and manufactured together in synthetic DNA. Together these trends make it possible to perform high-throughput experiments using synthetic DNA libraries in combination with screening experiments and NGS technology. The second is the decreasing cost of manufacturing synthetic DNA 10. The first trend is an increase in the number of short reads made available by Next Generation Sequencing (NGS) technology 9. But continuing trends in DNA reading and writing technologies are opening up new applications for large-scale DNA barcode libraries. Technological constraints have limited the size of the DNA barcode libraries used in screening experiments with the largest sized rationally designed DNA barcode library to date consisting of 240,000 barcodes 8. Randomly generated DNA libraries may be used for practical reasons such as cost, however, rationally designing DNA barcode libraries can have the advantage of being more robust against misidentification due to DNA sequencing and synthesis errors 7. DNA barcoding technology has shown utility in applications such as the discovery of new drug candidates 2 and elucidating protein interactions in yeast 6.ĭNA barcode libraries can be categorized into two groups, randomly generated DNA barcode libraries that are generated by physically assembling oligos in pools, and rationally designed DNA barcode libraries that are design in silico and then manufactured. In the past decade, libraries of DNA barcodes have found use in chemical compound screens 1, 2, the study of clonal diversity 3, and genomic screens 4, 5. Our results demonstrate the value of our novel large-scale DNA barcode library generation framework for use in high-throughput screening applications.Ī DNA barcode, or barcode for short, is a DNA sequence commonly used to identify a target molecule during DNA sequencing, but may also be used for other purposes such as the disruption of gene function or encoding information into a larger DNA region. We also report generating a general purpose one billion DNA barcode library, the largest such library yet reported in literature. As a proof of concept, we demonstrate that our framework is able to generate a library consisting of one million DNA barcodes for use in a fragment antibody phage display screening experiment. ![]() We show that our framework dramatically reduces the computation time required to generate large-scale DNA barcode libraries, compared with a naїve approach to DNA barcode library generation. Here we report a novel framework to quickly generate large-scale libraries of DNA barcodes for use in high-throughput screens. ![]() ![]() To be useful in experimental settings, the DNA barcodes in a library must satisfy certain constraints related to GC content, homopolymer length, Hamming distance, and blacklisted subsequences. In barcoded screens, DNA barcodes are linked to target biomolecules in a manner allowing for the target molecules making up a library to be identified by sequencing the DNA barcodes using Next Generation Sequencing. High-throughput screens allow for the identification of specific biomolecules with characteristics of interest. ![]()
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