3D printed integrated separator with hybrid micro-structures for high throughput and magnetic-free nucleic acid separation from organism samples

Abstract

• 3D-printed integrated separator for nucleic acid (NA) extraction was fabricated. • The separator has ideal printing accuracy and stable structures, and can be reused. • Hybrid micro-structures enhanced NA extraction efficiency of 3D-printed separator. • Developed magnetic-free NA extraction method is fast, simple and high-throughput. • Developed NA extraction method is promising in routine PCR diagnosis of pathogens. Nucleic acid (NA) amplification is a powerful molecular biological tool. While, the high-efficient NA amplification can be only used in modern laboratory owing to the relatively time-consuming and multistep labor-intensive NA separation. Herein, an integrated polyacrylic acid (PAA) comb-like separator with hybrid micro-structures was fabricated via 3D printing technology for high throughput and convenient (magnetic-free) NA separation. It realizes the high-effective and fast NA separation from organism samples by simply transferring of the separator from different tubes containing lysate, washing solution and elution buffer, with enhanced throughput due to the feature of convenient operation on 8-tube strip or 96-well plate. Results showed the 3D printing PAA integrated separator has ideal printing accuracy and stability in the thread and groove structures, and can be reused with robust separation performance. The NA separation yield enhanced by the hybrid micro-structures on the interface of the integrated separator is comparable to that of classic magnetic nanoparticle (MNP) based NA separation method under optimized condition. The developed NA separation and purification process has the advantages of very short operation time (separation duration minimized to 1 min), no requirement of additional technical equipment (magnetic-free) and no intensive labors (no pipetting and centrifugation). The NA amplification results illustrated that NA isolated via the developed separator could meet the requirements for further biological applications, especially in the field of high-throughput molecular detection of pathogens. This novel separator can provide a promising pave for NA separation to satisfy the high-throughput and automation requirements of the related biological applications.