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Hackflex: low cost Illumina sequencing library construction for high sample counts

View ORCID ProfileDaniela Gaio, View ORCID ProfileJoyce To, View ORCID ProfileMichael Liu, View ORCID ProfileLeigh Monahan, View ORCID ProfileKay Anantanawat, View ORCID ProfileAaron E. Darling
doi: https://doi.org/10.1101/779215
Daniela Gaio
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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Joyce To
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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Michael Liu
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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Leigh Monahan
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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Kay Anantanawat
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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Aaron E. Darling
1The ithree institute, University of Technology Sydney, Sydney, NSW, Australia
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  • For correspondence: aaron.darling@uts.edu.au
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ABSTRACT

We developed Hackflex, a low-cost method for the production of Illumina-compatible sequencing libraries that allows up to 11 times more libraries for high-throughput Illumina sequencing to be generated at a fixed cost. We call this new method Hackflex. Quality of library preparation was tested by constructing libraries from E. coli MG1655 genomic DNA using either Hackflex, standard Nextera Flex or a variation of standard Nextera Flex in which the bead-linked transposase is diluted prior to use. We demonstrated that Hackflex can produce high quality libraries and yields a highly uniform coverage, equivalent to the standard Nextera Flex kit. Using Hackflex, we were able to achieve a per sample reagent cost of library prep of A$8.66, which is 8.23 times lower than the Standard Nextera Flex protocol at advertised retail price. An additional simple modification to the protocol enables a further price reduction of up to 11 fold or about A$6.50/sample. This method will allow researchers to construct more libraries within a given budget, thereby yielding more data and facilitating research programs where sequencing large numbers of libraries is beneficial.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted September 23, 2019.
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Hackflex: low cost Illumina sequencing library construction for high sample counts
Daniela Gaio, Joyce To, Michael Liu, Leigh Monahan, Kay Anantanawat, Aaron E. Darling
bioRxiv 779215; doi: https://doi.org/10.1101/779215
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Hackflex: low cost Illumina sequencing library construction for high sample counts
Daniela Gaio, Joyce To, Michael Liu, Leigh Monahan, Kay Anantanawat, Aaron E. Darling
bioRxiv 779215; doi: https://doi.org/10.1101/779215

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