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Automated nucleic acid extraction from HepG2 cells application note

Automated Nucleic Acid Isolation from HepG2 cells

Nucleic Acid Extraction from HepG2 Cells: An Integrated Solution Utilizing the Bead Ruptor Elite™ Homogenizer, chemagic™ 360 Nucleic Acid Extractor and JANUS® G3 Automated Workstation

Applications featuring Next Generation Sequencing, quantitative PCR, or other genomics-based research using invitro cultured cells requires a nucleic acid extraction upstream of aforementioned methods. The Bead Ruptor Elite™ Bead Mill homogenizer and 2 mL bead kits are a streamlined sample preparation solution that provides efficiency in cell lysis, releasing nucleic acid for extraction on the chemagic™ 360 automated nucleic acid extractor.
In this aplication note, we outline the integration of the Bead Ruptor Elite™ Bead Mill homogenizer with the chemagic™ 360 nucleic acid extractor and Janus® G3 automated workstation, creating an end-to-end solution for sample preparation of in-vitro cultured cells and nucleic acid extraction workflows.

Automated nucleic acid extraction from HepG2 cells

Automated nucleic acid isolation from HepG2 cells

HepG2 DNA concentration from 12 samples

Lysate from samples 1-12 was prepared with Pro-K and RNase A. Average A260 /A280 from samples 1-12 was 2.21.

Nucleic Acid Extraction

After centrifugation step, 100 µl of cell lysate was transferred to 12 separate wells of a 96-well plate using the JANUS G3 automated workstation and customized Reagent Liquid Transfer Winprep program. Similarly, 100 µl Lysis Buffer provided with the chemagic Tissue DNA Kit (CMG-723) was transferred to each well containing the lysate. Next, 6 µl of Proteinase-K, included in the chemagic Tissue DNA kit, was manually transferred to all wells along with 5 µl RNase A. The plate was then incubated at 37°C for 10 minutes. During this time, Elution Buffer and Magnetic Beads were transferred into respective plates using a Reagent Liquid Transfer Protocol on the JANUS G3 automated workstation. After incubation, the tissue DNA extraction was carried out using the chemagic 360 nucleic acid extractor.

For research use only. Not for use in diagnostic procedures.