Development of 3D printed mesofluidic devices to elute and concentrate DNA

Cody Masters, Jocelyn Dolphin, April Maschmann, Keegan McGill, Matthew Moore, Drew Thompson, Kristy L. Kounovsky-Shafer

Research output: Contribution to journalArticle

Abstract

To understand structural variation for personal genomics, an extensive ensemble of large DNA molecules will be required to span large structural variations. Nanocoding, a whole-genome analysis platform, can analyze large DNA molecules for the construction of physical restriction maps of entire genomes. However, handling of large DNA is difficult and a system is needed to concentrate large DNA molecules, while keeping the molecules intact. Insert technology was developed to protect large DNA molecules during routine cell lysis and molecular biology techniques. However, eluting and concentrating DNA molecules has been difficult in the past. Utilizing 3D printed mesofluidic device, a proof of principle system was developed to elute and concentrate lambda DNA molecules at the interface between a solution and a poly-acrylamide roadblock. The matrix allowed buffer solution to move through the pores in the matrix; however, it slowed down the progression of DNA in the matrix, since the molecules were so large and the pore size was small. Using fluorescence intensity of the insert, 84% of DNA was eluted from the insert and 45% of DNA was recovered in solution from the eluted DNA. DNA recovered was digested with a restriction enzyme to determine that the DNA molecules remained full length during the elution and concentration of DNA.

Original languageEnglish (US)
Pages (from-to)810-816
Number of pages7
JournalELECTROPHORESIS
Volume40
Issue number5
DOIs
StatePublished - Mar 2019

Keywords

  • 3D printing
  • DNA concentration device
  • DNA elution

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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  • Cite this

    Masters, C., Dolphin, J., Maschmann, A., McGill, K., Moore, M., Thompson, D., & Kounovsky-Shafer, K. L. (2019). Development of 3D printed mesofluidic devices to elute and concentrate DNA. ELECTROPHORESIS, 40(5), 810-816. https://doi.org/10.1002/elps.201800309