BioCloneBot: A versatile, low-cost, and open-source automated liquid handler

Abstract

Liquid handler systems can provide significant benefits to researchers by automating laboratory work, however, their unaffordable price provides a steep barrier to entry. Therefore, we provide the BioCloneBot, a versatile, low-cost, and open-source automated liquid handler. This system can be easily built with 3D-printed parts and readily available commercial components. The BioCloneBot is highly adaptive to user needs and facilitates various liquid handling tasks in research and diagnostics. Its user-friendly interface and programmable nature make it suitable for a wide range of applications, from small-scale experiments to larger laboratory setups. By utilizing BioCloneBot, researchers and scientists can streamline their liquid handling processes without the financial constraints posed by traditional systems. In this paper, we detail the design, construction, and validation of BioCloneBot, showcasing its precise control, accuracy, and repeatability in various liquid handling tasks. The open-source nature of the system encourages collaboration and customization, enabling researchers to contribute and adapt the technology to specific experimental requirements.

Keywords: 3D printing; DNA cloning; Liquid handler; Open-source; Synthetic biology.

Graphical abstract

Fig. 1

Custom Microliter Syringe Pump with P200 Tip Adapter.

Fig. 2

BioCloneBot frame and pump fully assembled.

Fig. 3

3D model of platform where labware is placed and experiments are performed.

Fig. 4

BioCloneBot Front-End.

Fig. 5

The left side of the front-end includes the Protocol Queue which displays the selected operations in order. Also included are the potential Operations and Available Labware. Underneath the Protocol Queue are buttons used to control the software such as Start Experiment, Clear Protocol, Load Sample Experiment, and more.

Fig. 6

The right side of the front-end shows the platform for placing labware.

Fig. 7

The properties window supports the selection of individual wells, tubes, or tips. The select all button allows for quick modification of reservoir volumes or tip availability. Modifications are only confirmed once the OK button is pressed.

Fig. 8

BCB raphical User Interface for Lab Experiment.

Fig. 9

Restriction enzyme digest of CEN6/URA3 Kan+ plasmid with GFP dropout insert. Digestion with restriction enzyme BsaI results in a separation of the backbone at 3721 bp and the GFP insert at 1026 bp. The full plasmid length is 4747 bp.

Fig. 10

Gibson Assembly of CEN6/URA3 GFP dropout plasmid using automated liquid han- dler. Reaction was done under the Hifi Assembly protocol where each fragment, GFP and CEN6/URA3 Backbone were PCR amplified with specific 20 nucleotide overhangs.

Fig. 11

Golden Gate Cloning assembly of mRuby construct using the BioCloneBot. Diges- tion and ligation using the NEB Golden Gate assembly kit and plasmid: pYTK011 (pPGK1), pYTK034 (mRuby), pYTK056 (tTDH1) and CEN6/URA3-GFP dropout backbone.

Fig. 12

Miniprep of isolated colonies from automated and manual assembly. The full length of assembly is 5733 bp. Digestion with NotI results in bands at 3614bp and 1763 bp.