Setting up a 384 well qRT-PCR assay for the ViiA™ 7 using VIAFLO and VOYAGER electronic pipettes

  • Setting up a 384 well qRT-PCR assay for the ViiA™ 7 using VIAFLO and VOYAGER electronic pipettes

    Pipetting 64 samples in triplicate for two genes of interest (GOI), all under 10 minutes!

    The qRT-PCR is set up by hand, using a VIAFLO 16 channel 125 μl electronic pipette to distribute master mixes of two genes of interest into a 384 well plate. All 64 samples are loaded directly from 1.5 ml microcentrifuge tubes into the 384 well plate in triplicate, using a VOYAGER 8 channel 12.5 μl adjustable tip spacing pipette. The whole process takes less than 10 minutes in real time!

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      The qRT-PCR is set up by hand, using a VIAFLO 16 channel 125 μl electronic pipette to distribute master mixes of two genes of interest into a 384 well plate. All 64 samples are loaded directly from 1.5 ml microcentrifuge tubes into the 384 well plate in triplicate, using a VOYAGER 8 channel 12.5 μl adjustable tip spacing pipette. The whole process takes less than 10 minutes in real time!

    Key benefits

    • A 384 well plate can be filled with a master mix in less than one minute, using a VIAFLO 16 channel pipette and the Repeat Dispense mode.
    • The VOYAGER adjustable tip spacing pipette enables simultaneous transfer of multiple samples from microcentrifuge tubes to a 384 well plate, saving many extra pipetting steps and time.
    • Reducing the number of individual transfers minimizes the chance of pipetting errors.

    Set up a 384 well qRT-PCR plate by pipetting 64 samples in triplicate

    The video shows how quickly and easily you can set up a 384 well qRT-PCR plate by pipetting 64 samples in triplicate for two genes of interest (GOI 1 and GOI 2).

    Both master mixes are transferred from a microcentrifuge tube into a 10 ml polystyrene multichannel reagent reservoir using a single channel pipette. The GOI 1 master mix is then transferred to the left half of the 384 well plate, and the GOI 2 master mix to the right half, using the Repeat Dispense mode of the VIAFLO 16 channel 125 μl electronic pipette. Next, all 64 samples are transferred in triplicate into both the GOI 1 and GOI 2 master mixes using a VOYAGER 8 channel 12.5 μl pipette. The VOYAGER pipette’s tip spacing can be adjusted on the fly for center-to-center spacing of the tubes and the plate, enabling simultaneous direct transfer of multiple samples.

    Video: Setting up a 384 well qRT-PCR assay using INTEGRA’s electronic pipettes

    Step-by-step procedure

     

    1. Transfer GOI 1 and GOI 2 master mixes into a 384 well plate 

    Add GOI 1 master mix to the left half of a 384 well plate and GOI 2 master mix to the right half (see Figure 1).

    Use a VIAFLO 16 channel 125 μl electronic pipette with 125 μl Sterile, Filter GripTips.
    Select the Repeat Dispense mode and set it to dispense 12 x 7.5 μl. Depending on the viscosity of the master mix, a lower pipetting speed (e.g. speed 6) is recommended. A Pace option can be activated to minimize the need to press the pipette’s run button during dispensing, and enable automatic sequential dispensing.

    Figure 1: Pipetting scheme for GOI 1 and GOI 2 master mixes
    Figure 1: Pipetting scheme for GOI 1 and GOI 2 master mixes

    2. Transferring 64 samples into a 384 well plate

    Add 64 samples of GOI 1 and GOI 2 in triplicate to the master mix.

    All 64 samples in microcentrifuge tubes are divided into 2 x 32 samples lined up in either a yellow or green rack and then transferred in triplicate to the GOI 1 and GOI 2 master mixes (see Figure 2), using a VOYAGER 8 channel 12.5 μl adjustable tip spacing pipette with 12.5 μl Sterile, Filter GripTips.
    Choose the Repeat Dispense mode and set it to dispense 3 x 2.5 μl. A post-dispense of 1-2 μl is recommended for optimal dispensing performance. Two tip spacings are set: 13 mm (tubes in the tube rack) and 4.5 mm (center-to-center spacing of 384 well plates).

    Figure 2: Pipetting scheme of the qRT-PCR assay
    Figure 2: Pipetting scheme of the qRT-PCR assay

    December 2019, by

    • Lydiane Saucède, PhD

      Application Scientist

      Email

    Any questions? I'm happy to help!

    • Tom Bentivegna

      Product Manager

      Email

    Used instruments and accessories

    VOYAGER, 8 Channel, 12.5 µl VIAFLO 16 channel, 125 µl, electronic pipette Multichannel Reagent Reservoir, 10 ml, PP GripTip, 12.5 µl, Sterile, Filter GripTip, 125 µl, Sterile, Filter

    VOYAGER, 8 Channel, 12.5 µl

    VOYAGER pipettes allow the tip spacing to expand anywhere between 4.5 mm and 33 mm at the push of a button.

    • Single handed operation leaves the other hand free to handle labware.
    • On the fly access to up to 3 user-defined tip spacings.
    • These user defined tip spacings are saved. No need to memorize spacings of different labware formats.

    Part No. 4721

    See it work
    VOYAGER adjustable tip spacing pipette

    VIAFLO 16 channel, 125 µl, electronic pipette

    The Touch Wheel is a quick and ergonomic way to modify pipetting parameters. Instead of repeatedly pushing buttons or twisting fingers to modify volumes, you simply slide your finger over the wheel.

    The full color screen provides full text menus (in multiple languages) and displays pipetting protocols without abbreviations, making VIAFLO pipettes particularly easy to understand and intuitive to use.

    Part No. 4642

    Find out more about VIAFLO electronic pipettes
    INTEGRA’s range of VIAFLO electronic pipettes.

    Multichannel Reagent Reservoir, 10 ml, PP

    • SureFlo™ anti-sealing array prevents pipette tip seal off which can cause liquid to pop into the tip, filter or even the pipette itself
    • Unique surface treatment spreads liquid evenly offering even lower dead volumes
    • Reusable base with bold, crisp, clearly visible graduation markings
    • Nest inside each other, making it possible to get twice as many reservoirs in half the space of other products on the market
    • Reducing inventory space requirements and shipping costs
    • Available as crystal clear polystyrene reservoirs or polypropylene reservoirs for improved chemical compatibility
    Find out more about Multichannel Reagent Reservoirs
    Reagent reservoirs 10ml, 25ml, 100ml

    GripTip, 12.5 µl, Sterile, Filter

    GripTip pipette tips perfectly match the multi-lobe tip fitting, snapping firmly on during loading. This guarantees a perfect seal on every tip, preventing them from loosening, leaking or completely falling off. All tips are precisely aligned horizontally, enabling accurate touch-offs, even when pipetting with 384 tips.

    Part No. 4415

    Learn more about GripTips pipette tips for Benchtop Pipetting Systems
    Integra-Relaunch_Website-Output-Produktebilder-Product_Overview-GripTips.png

    GripTip, 125 µl, Sterile, Filter

    GripTip pipette tips perfectly match the multi-lobe tip fitting, snapping firmly on during loading. This guarantees a perfect seal on every tip, preventing them from loosening, leaking or completely falling off. All tips are precisely aligned horizontally, enabling accurate touch-offs, even when pipetting with 384 tips.

    Part No. 4425

    Learn more about GripTips pipette tips for Benchtop Pipetting Systems
    Integra-Relaunch_Website-Output-Produktebilder-Product_Overview-GripTips.png

    Customer’s voice

    There are clear advantages of manual set-up using VIAFLO and VOYAGER pipettes. While liquid handling robots can be a good option for routine processing of large sample numbers, handheld pipettes like these give far greater control over changing sample numbers and layouts. You can pipette a 384 well plate by hand in the time it takes to set up the robot.