Goal
The Thermo Scientific™ KingFisher™ Duo Prime automated purification system offers a fast way to extract high quality nucleic acids in low-to medium-throughput labs. With the Thermo Scientific™ E1-ClipTip™ Adjustable Tip Spacing Equalizer Multi-channel Electronic Pipette it is possible to enhance the extraction process by using only one tool to do all of the pipetting – adding samples, reagents and magnetic beads on the deep-well 96-plate or 12-well elution strip can be done entirely using one pipette.
The E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette will save valuable lab time and reduce the risk of incurring injuries related to the repetitive motions required in the process.
Introduction
The samples for DNA extraction with the KingFisher Duo Prime purification system may vary from plant leaves to mouse ears, or any organism of interest. The sample can be extracted using a variety of sample vessels available; e.g. blood tubes of different sizes, 15 mL/50 mL conical tubes, microcentrifuge tubes or microplate formats.
The KingFisher Duo Prime purification system works in standard 24- and 96-plate formats and typically there is a need to use several pipettes of different volume settings to fill the reagents on the plate and the elution strip. However, the need to switch between pipettes has been removed.
Thermo Scientific KingFisher deepwell 96-plate and KingFisher Duo Prime elution strips can easily be filled using the E1-ClipTip Equalizer pipette. The adjustable tip spacing feature makes it possible to transfer samples from tubes to plates, and dispense magnetic particles and buffers effortlessly with multi-dispensing pipetting functionality. It is also easy to program, save and name a ready-to-use protocol to fill a plate for routine applications in the Programs mode. E1-ClipTip pipettes feature a unique Matrix function that utilizes step-based programming which allows the creation of fully customized pipetting protocols. As such, this provides an ideal option for users that require a variety of pipetting functions in a unique or specific order to complement different research protocols.
In this article, the simplicity of using the E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette to dispense samples and reagents on the King-Fisher Duo Prime is described with an example of DNA extraction from human cells. One pipette is used for all sample and reagent steps with easy access to useful functions such as mixing and multi-dispensing, to increase pipetting efficiency.
Materials and Methods
The E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette was programmed for pipetting all samples and reagents for DNA purification from human cells using the KingFisher Cell and Tissue DNA Kit on the KingFisher Duo Prime purification system. 12 parallel samples of 5 x 105 HeLa cells were used for testing. Cell lysis was performed according to kit instructions and the lysed samples were pipetted
from microcentrifuge tubes to the KingFisher deep-well 96-plate using a pre-programmed E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette.
Two programs were generated using the Matrix function. One program was created to guide users through the sample row pipetting
“KF CT Sample row Duo” and another one for pipetting washing and elution buffers “KF CT Buffers Duo.” Shortcuts can be created for the most commonly used programs on the E1- ClipTip main menu for straightforward and quick access in the laboratory.
KingFisher Cell and Tissue DNA Kit instructions were followed to adjust steps and volumes on the E1-ClipTip program according to Table 1. Thermo Scientific ClipTip 1250 pipette tips were used in all pipetting steps. The samples and reagents were pipetted on the KingFisher deep-well 96 plate except for the Elution Buffer, which was pipetted on the KingFisher Duo elution strip.
In order to achieve uniform results, it is extremely important that a homogenous solution of magnetic beads is reached. To ensure this evenness of bead distribution, the magnetic beads are mixed using the pipette, before aspiration of the solution for multiple dispense using a single channel of the E1-ClipTip pipette.
The “KF_TissueDNA_Duo” protocol was started on the KingFisher Duo Prime purification system to perform the DNA purification. Once filled, the deep-well 96-plate and 12-well elution strip were inserted into the instrument following the instructions on the user interface. The Thermo Scientific™ Multiskan™ GO microplate spectrophotometer was then used to determine DNA quantity and quality by common absorbance measurements.
Results
Genomic DNA (gDNA) purification from HeLa cells resulted in even yields of high quality gDNA. The resulting agarose gel shows clear bands indicating intact gDNA, and the absorbance spectra shows extracted gDNA to be pure without remains of proteins or other impurities (Figures 1a and 1b).
Conclusions
The E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette with adjustable tip spacing is an ideal solution for dispensing samples and reagents for nucleic acid purification using the KingFisher Duo Prime purification system. By using the E1-ClipTip Equalizer 6-channel pipette, it is possible to use just one pipette and one size of pipette tip to easily dispense all reagents across all protocol steps, when dispensing volumes from 25 to 800 µL. The adjustable tip spacing helps to reduce repetitive motions, thus increasing personnel safety and lab ergonomics, while reducing pipetting time when working between different labware formats. As a result, laboratory efficiency is increased.
About the Author
Sini Suomalainen is an Applications Manager with Thermo Fisher Scientific.
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