BenchSmart 96. Semi-automated Pipetting Higher Accuracy, Greater Flexibility

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Kerry Horton
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BenchSmart 96 Semi-automated Pipetting Higher Accuracy, Greater Flexibility For scientists looking to maximize their data quality and research productivity, a new semiautomated approach to 96-well

1 BenchSmart 96 Semi-automated Pipetting Higher Accuracy, Greater Flexibility For scientists looking to maximize their data quality and research productivity, a new semiautomated approach to 96-well pipetting is a compelling option. Neither entirely manual nor wholly robotic, in a research environment semi-automation can deliver superior accuracy and repeatability in a highly flexible format. As a fundamental activity in biological laboratory research science, pipetting can be a timeconsuming and repetitive task. In most bioscience laboratories in the world, manual pipettes are used in both single and multichannel formats to fill receptacles such as microfuge tubes, microtiter plates, and more. In terms of high-throughput experimental workflows, many scientists still rely on manual multichannel pipettes and 96- or 384-well plates to carry out their work. While multichannel pipettes definitely accelerate plate-based experiments, additional options such as 96-channel pipetting stations and robotics are available, albeit at a higher price point than smaller handheld manual devices. Though more expensive, the time savings for 96-channel pipetting stations and robotics over multichannel pipettes is worth the expense to many scientists who need to maximize their data quality and research productivity. has offered the manual 96-channel Liquidator 96 since 2010 recognizing that operational simplicity matters almost as much as pipetting accuracy and precision. The power of the manual Liquidator 96 system lies in the fact that no programming knowledge is necessary to operate the device and the device carries out simple up-and-down pipetting for 96- and 384- plate applications. Electronic pipetting can also be complimentary to manual pipetting. Sometimes, liquid handling workflows are so complicated that for manual systems, convenience is sacrificed to finish a workflow in a timely manner. Additionally, even 96-channel devices can cause a user's hand to eventually tire, which can make accurate and precise pipetting a challenge. For this reason, introduced the BenchSmart 96 pipetting station to facilitate semiautomated liquid handling in 96- and 384-well plates. The BenchSmart 96 offers the benefits of manual 96-channel pipetting, but adds the benefits of electronic pipetting, which can be used in multiple modes such as basic, multidispense, advanced pipetting, and dilute mode to carry out sophisticated liquid handling protocols.

BenchSmart 96 Benefits of the BenchSmart 96 include: Multiple volume ranges three heads to accommodate pipetting from 0.5 to 1,000 µl. Convenience interchangeable heads allow for bench-space savings.

2 BenchSmart 96 Benefits of the BenchSmart 96 include: Multiple volume ranges three heads to accommodate pipetting from 0.5 to 1,000 µl. Convenience interchangeable heads allow for bench-space savings. Accuracy and precision the BenchSmart 96 exhibits excellent accuracy and precision for sensitive applications in genomics and proteomics. Reproducible the BenchSmart's 96 channels aspirate and dispense consistently. Tested the BenchSmart 96 have been shown to function well in ELISA and qpcr. Fewer errors unlike single and multichannel pipettes, the BenchSmart 96 pipettes all 96-channels simultaneously, eliminating the possibility of skipping or repeating rows and wells. Simple operation the touchscreen user interface on the BenchSmart 96 is easily navigated, and can be used to change pipetting settings in an intuitive manner. Ergonomic the effort of manual pipetting is eliminated with the BenchSmart 96, and the large touchscreen is highly visible. Electronic functionality The BenchSmart can be used in multiple electronic pipetting modes including basic, multidispense, advanced pipetting, and dilute. Inexpensive The BenchSmart 96 is well within reach for both academic and industrial customers. The BenchSmart's numerous benefits make it a formidable tool for biological and chemical research laboratories. This document describes how BenchSmart 96 can be used to carry out common protocols in biological laboratories. Figure 1. BenchSmart 96 2

3 Performance of BenchSmart 96 in qpcr Quantitative polymerase chain reaction (qpcr) is perhaps the most sensitive laboratory application in terms of pipetting precision. Especially for template pipetting steps, differences in volumes pipetted can result in changes for observed Cq due to different amounts of amplifiable material present in the reaction mixture. The BenchSmart 96 with 20 µl head was compared to a single channel 20 µl electronic pipette as well as a eight channel 20 µl electronic pipette in carrying out pipetting for qpcr (Figure 2A, 2B, and 2C). In this experiment, each pipetting device was used to pipette 18 µl of qpcr mastermix into a 96-well plate prior to pipetting 2 µl of DNA template at 10 ng/µl into each well of the same plate. The pipetting steps were carried out in basic pipetting mode, with blowout on for all steps. After the second pipetting step, the plate was sealed, vortexed, then placed in a qpcr machine and analyzed. The data obtained shows that in this particular experiment, the BenchSmart 96 provided data that is within error to that obtained by single and 8-channel electronic pipettes. The uncertainty for these measurements was also comparable. To check whether changing pipetting parameters would affect data quality, the BenchSmart 96 was used to pipette 18 µl of mastermix into a 96-well plate, followed by 2 µl of 1 ng/µl DNA template. For each of these steps, advanced mode was used with the BenchSmart 96 and the blowout was turned off for each step. The data shown below had excellent reproducibility as evident in small uncertainty observed in the mean Cq measurement (Figure 2D). A Amplification RFU Cycles B Amplification RFU Cycles 3

4 BenchSmart 96 C RFU Amplification Cycles Amplification D RFU Cycles Figure 2. BenchSmart 96, single, and multichannel electronic pipette performance in qpcr. Panel A. A single channel 20 µl electronic pipette was used to set up qpcr in two pipetting steps involving an initial 18 µl mastermix addition step followed by a 2 µl addition of 10 ng/µl template DNA. Blowout was on for all steps. The plate was sealed, vortexed, then analyzed by qpcr. The mean Cq value across an entire 96-well plate was ± Panel B shows analogous data, with the exception that a eight channel pipette was used for all pipetting steps. The mean Cq value across an entire 96-well plate was ± Panel C shows the same data with the exception that the BenchSmart 96 was used for pipetting. The mean Cq value across an entire 96-well plate was ± Panel D shows data from a qpcr experiment where the BenchSmart 96 was used for pipetting mastermix and 1 ng/µl DNA template. In this experiment the BenchSmart was used in advanced pipetting mode with all reagents. The blowout was left off for all steps. The mean Cq value across an entire 96-well plate was ± Taken together, the data above shows that the BenchSmart 96 functions analogously to single and multichannel electronic pipettes, while providing excellent data. 4

5 The BenchSmart 96 with a 200 µl head was also tested in a proteomics workflow. The BenchSmart was used to generate eight replicate standard curve data sets in quantitative ELISA. After setting up initial sample dilutions with an electronic 8-channel pipette, the BenchSmart was used to carry out all subsequent steps of the ELISA protocol depicted below. The BenchSmart was able to generate standard curves with good linearity, comparable to the Liquidator 96 device (Figure 3). This data shows that the BenchSmart 96 can provide data similar to the Liquidator 96 in proteomics applications. A B Figure 3. Comparison of Liquidator 96 and BenchSmart 96 in ELISA Standard Curve Generation. Panel A shows the average of 8 standard curves generated with the Liquidator 96, while Panel B shows the average of 8 standard curves generated with the BenchSmart 96. 5

6 BenchSmart 96 Finally, the BenchSmart 96 with 200 µl head was compared against another manufacturer's 96-channel pipetting device in carrying out multidispense pipetting. The BenchSmart 96 was analyzed in pipetting 20 % (40 µl) of the 200 µl head's total volume. To analyze the amount of liquid pipetted, the BenchSmart was used to aspirate 200 µl of 100 µm Cibacron Blue dye dissolved in 50 mm Tris-HCl (ph 7.6). After multidispensing into multiple plates, the plates were then read at 615 nm in a platereader. This workflow was also carried out to analyze the ability of the other manufacturer s 96-channel pipetting device in order to gain an understanding of which device provides superior data. The average absorbance data for all plates for the BenchSmart 96 was ± 0.008, while it was ± for the other device. In terms of intra-plate consistency, the BenchSmart repeatedly gave lower standard deviations per plate versus the other manufacturer s device. BenchSmart 96 Data Dispense Number Mean Absorbance Standard Deviation Total Plates Other Manufacturer s Data Dispense Number Mean Absorbance Standard Deviation Total Plates

7 Perspective The BenchSmart 96 is a semi-automated manual pipetting system with electronic functionality that is designed for 96- and 384-well plate assays. Two such assays are qpcr and ELISA which are used to detect and quantify nucleic acids and antigen proteins, respectively. Quantitative PCR is highly sensitive to small differences in template nucleic acid concentrations. If a pipette adding template nucleic acid to a qpcr mixture is imprecise, then the imprecision will be evident in high variation in Cq values along a 96-well qpcr plate. Single channel and multichannel electronic pipettes have been used with success in qpcr, so comparison of the BenchSmart 96 with these devices would provide an indication of the pipetting capability/quality of the BenchSmart 96. The BenchSmart, like single and multichannel electronic pipettes, is able to provide high quality qpcr data, as evident in the data shown in this document. ELISA can be carried out qualitatively or quantitatively, and in the latter case, the accuracy and precision by which an ELISA standard curve is generated can be critical. Multichannel pipettes cannot add buffer solutions and protein samples to all wells in 96-well plates simultaneously. For this reason, washing and incubation times might diverge for different samples in the same plate and the final standard curve might not be of high quality. The BenchSmart, like the Liquidator 96, is able to provide high quality data in ELISA. Key benefits that the BenchSmart 96 offers in qpcr and ELISA are: The ability to add buffer and samples to all wells in 96 wells of a plate simultaneously Improved linearity and extended range of standard curves, which allows for the calculation of more accurate protein concentrations, with the reduced need of diluting samples so that their absorbance values fall within standard values. There is no possibility of skipping wells in a 96-well plate, which could compromise the data from an otherwise sound experiment. Increased throughput and data quality are important for modern biological researchers. The faster samples can be processed and high quality data obtained, the more rapid valid conclusions can be reached and then published. The BenchSmart 96 is able to speed customer workflows with features such as multidispense pipetting. Compared to pipettes with less channels, the BenchSmart 96 can deliver both speed and high data quality to qpcr, ELISA and other assays. 7

8 Mettler-Toledo, LLC 7500 Edgewater Drive, Oakland, CA Phone Fax For more information Subject to technical changes 03/2016 Mettler-Toledo, LLC Printed on demand from online content. WP-516 Rev A (A4)

Liquidator 96. CCDB Simplifies Genomic Workflow Liquidator 96 Used in DNA Barcoding

Liquidator 96. CCDB Simplifies Genomic Workflow Liquidator 96 Used in DNA Barcoding CCDB Simplifies Genomic Workflow Liquidator 96 Used in DNA Barcoding The Canadian Centre for DNA Barcoding (CCDB) uses the throughout its barcoding workflow. The CCDB is developing a barcode reference