Nucleic acid extraction with the Arcis Sample Prep Kit

  • Nucleic acid extraction with the Arcis Sample Prep Kit

    Automatic nucleic acid extraction with the ASSIST PLUS pipetting robot in just two steps!

    The Arcis Sample Prep Kit is a ready-to-use kit comprising two reagents that enables pre-analytical processing of a variety of biological sample types, including whole blood, saliva, bacteria and plant cells. DNA/RNA extraction can be so simple; the Arcis Sample Prep Kit allows the user to go from cells to downstream nucleic acid investigations in only three minutes, without the need for isolation or purification. The protocol consists of just two steps that can be easily automated with the ASSIST PLUS pipetting robot, offering the user a convenient solution for nucleic acid extraction. Sterile working conditions are essential, especially for RNA extractions. The ASSIST PLUS meets the requirements for working under sterile conditions, as its small footprint means that it can be easily placed in a laminar flow cabinet.

  • Table of contents

      The Arcis Sample Prep Kit is a ready-to-use kit comprising two reagents that enables pre-analytical processing of a variety of biological sample types, including whole blood, saliva, bacteria and plant cells. DNA/RNA extraction can be so simple; the Arcis Sample Prep Kit allows the user to go from cells to downstream nucleic acid investigations in only three minutes, without the need for isolation or purification. The protocol consists of just two steps that can be easily automated with the ASSIST PLUS pipetting robot, offering the user a convenient solution for nucleic acid extraction. Sterile working conditions are essential, especially for RNA extractions. The ASSIST PLUS meets the requirements for working under sterile conditions, as its small footprint means that it can be easily placed in a laminar flow cabinet.

    Key benefits

    • Automating the Arcis Sample Prep Kit eliminates the risk of cross-contamination between samples and ensures a consistent workflow.
    • The ASSIST PLUS guides the user through the protocol, making the pipetting process less error-prone and the results more reproducible.
    • VIAFLO and VOYAGER electronic pipettes, in combination with ASSIST PLUS, provide unmatched pipetting ergonomics.
    • The universal deck of the ASSIST PLUS allows easy adjustment and positioning of all kinds of individual labware, including blood test tube racks.

    Overview: Nucleic acid extraction

    Note: Make sure that all samples have thawed completely before starting this procedure!

    In this protocol, DNA is extracted from whole blood samples using the Arcis Sample Prep Kit and the ASSIST PLUS. The pipetting robot operates a VOYAGER 8 channel 125 μl electronic pipette with 125 μl Sterile, Filter, Low Retention GripTips. The use of Low Retention GripTips ensures optimal handling of viscous liquids, such as blood samples.

    The protocol is divided into two parts:

    • Program 1: Blood sample transfer (ARCIS_Transfer_WholeBlood)
    • Program 2: Nucleic acid extraction with the Arcis Sample Prep Kit (ARCIS_DNA_Extraction)
    Arcis Sample Prep Kit (50 rxn) consists of Lysis Buffer and Wash Buffer.

    Downloads: Protocols for automated nucleic acid extraction with ASSIST PLUS

    Here you can download the complete protocols and reports used in this application and use them on your ASSIST PLUS. The protocol can be customized with the VIALAB software.

    Experimental set-up: Program 1

    Deck position B:

    96 deep well plate - empty.

    Deck position C:

    Rack for 6 ml EDTA blood test tubes (12 x 100 mm) – 6000 μl.

    Set-up for blood sample transfer
    Figure 2: Set-up for blood sample transfer

    Step-by-step procedure

    1. Aliquot the blood samples

    Transfer the blood samples from the collection tubes.

    Place the blood collection tubes on position C and a 700 μl deep well plate on position B. Then select the ARCIS_SAMPLES program on the VOYAGER electronic pipette by pressing Run. The ASSIST PLUS immediately starts to transfer blood from the collection tubes into the deep well plate.

    Tip: Use the Tip Travel feature of the ASSIST PLUS to keep the tip immersion depth constant during aspiration and dispensing, ensuring contamination-free and consistent liquid handling for more reproducible results, no matter how often the application is repeated.

    Experimental set-up: Program 2

    Deck position A:

    Dual Reservoir adapter with 2 x 10 ml multichannel reagent reservoirs for Reagent 1 (blue) and Reagent 2 (green).

    Deck position B:

    96 deep well plate - partially filled with 100 μl of blood samples (pink).

    Deck position C:

    96 well PCR Sapphire - empty.

    Set-up for Arcis DNA extraction
    Figure 3: Set-up for Arcis DNA extraction

    Step-by-step procedure

    1. Reagent distribution

    Aliquot Reagent 1 and Reagent 2.

    Place two 10 ml multichannel reagent reservoirs into a dual reservoir adapter from INTEGRA on position A, the deep well plate with the preloaded blood samples on position B, and a new 96 well plate on position C. Fill the two 10 ml multichannel reagent reservoirs with Reagent 1 and 2, and change the program on the VOYAGER pipette to ARCIS_KIT. The ASSIST PLUS then starts to aliquot 150 μl of Reagent 1 in to the deep well plate (position B, columns 7-12) followed by the distribution of 20 μl of Reagent 2 in the 96 well plate (position C, columns 1-6).

    Tips:

    • A pre- and post-dispense step is recommended to increase the accuracy and precision of pipetting. The pre- and post-dispense volume should be between 3 and 5 % of the nominal volume of the pipette.
    • Save reagents by using the 10 ml multichannel reagent reservoir with SureFlo anti-sealing array. This allows you to aspirate from the very bottom of the reservoir in order to minimize the dead volume of the reagents.

    2. Sample lysis

    Add blood samples to Arcis Reagent 1.

    Transfer 90 μl of each blood sample to the wells containing Reagent 1. After mixing, there is a one-minute incubation period before the samples are further processed or stored for processing later, according to the manufacturer’s protocol (Arcis Sample Prep Kit).

    Tip: Be sure to mix Reagent 1 thoroughly with the blood samples to ensure a homogenous solution.
    Note: Nucleic acids are stable for 90 days at room temperature, provided there is no further processing.

    3. Sample washing

    Wash lysed samples with Arcis Reagent 2.

    Continuing this protocol, use the ASSIST PLUS to automatically transfer 5 μl of the lysed blood samples (position B) to Reagent 2 in the new 96 well plate (position C) and mix them thoroughly.

    Tip: After addition of Reagent 2, the samples must be used immediately or frozen at -20 ºC. To reduce sample dilution, samples can be added to Reagent 2 at a ratio of 1:3, 1:2 or 1:1.

    Remarks

    • The PCR reaction can be run immediately or go directly to other downstream techniques. To continue with a PCR reaction, add the appropriate volume to the PCR master mix (e.g. 5 μl per 25 μl of PCR reaction).
    • Users who only want to extract DNA from eight samples can place an 8 well PCR strip in a PCR 96 well cooling block (INTEGRA) instead of a 96 well plate.
    • To process a higher number of samples consider using the kit for 500 reactions and choose the VIAFLO 96/384 to add reagents or samples simultaneously.

    Conclusion

    • Rapid nucleic acid extraction without the need to purify or isolate blood samples.
    • VIALAB software can easily be adjusted to the labware required by the laboratory at any time.
    • Automation of the workflow eliminates the risk of human error and increases the reproducibility of the results.
    • The easy implementation of the Arcis Sample Prep Kit with the ASSIST PLUS pipetting robot accelerates pre-analytical processing in human and microbiological samples.

    April 2020, by

    • Lydiane Saucède, PhD

      Application Scientist

      Email
    • Jan-Peter Baldin, PhD

      Product Manager

      Email

    Any questions? I'm happy to help!

    • Stephanie Stursberg

      Product Manager

      Email

    Used instruments and accessories

    ASSIST PLUS, Pipetting Robot VOYAGER, 8 Channel, 125 µl GripTip, 125 µl, Sterile, Filter, Low retention Dual Reservoir Adapter Multichannel Reagent Reservoir, 10 ml, PP Rack for 5 ml test tubes, 6x8 tubes PCR 96 well cooling block CORNING: Corning® Thermowell ® GOLD 0.2 ml 8-Well PCR tube strips Arcis Sample Prep Kit 50rxn Eppendorf, 500 µl Deepwell Plate

    ASSIST PLUS, Pipetting Robot

    INTEGRA has developed the ASSIST PLUS pipetting robot to streamline routine pipetting tasks at an affordable price. Using INTEGRA electronic multichannel pipettes, the system:

    • automates pipetting tasks,
    • eliminates physical strain and
    • ensures superior reproducibility and
    • error free pipetting.

    Part No. 4505

    Visit product page
    ASSIST PLUS Pipetting Robot

    VOYAGER, 8 Channel, 125 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. 4722

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

    GripTip, 125 µl, Sterile, Filter, Low retention

    INTEGRA uses a unique polypropylene blend to mold Low Retention GripTips with heightened hydrophobic properties. Using Low Retention GripTips prevents low surface tension samples from spreading out and “wetting” the inner wall of the tips, allowing them to bead-up for a maximum liquid recovery.

    Part No. 6565

    Learn more about low retention pipette tips
     Low Retention GripTips are offered in 6000 series racks as non-sterile, sterile and filter options.

    Dual Reservoir Adapter

    The dual reservoir adapter makes this system ideal for assays that required sequential addition of several low volume reagent mixes. This adapter holds two 10 ml, 25 ml or divided reagent reservoirs side by side in an aluminum frame, and can be pre-chilled to keep reagents cool during processing.

    Part No. 4547

    Find out more about ASSIST PLUS accessories
    Dual reservoir adapter with 2 didived reservoirs

    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

    Rack for 5 ml test tubes, 6x8 tubes

    Rack for 5 ml test tubes (12x75 mm type), 6x8 tubes

    Part No. 4543

    Find out more about ASSIST PLUS accessories
    Tube Racks for Centrifuge Tubes, Test Tubes and Cryogenic Tubes.

    PCR 96 well cooling block

    Aluminium 96 and 384 well PCR cooling blocks that keep reagents chilled during sample preparation in a PCR plate. The cooling blocks feature a SBS footprint.

    Part No. 6250

    Find more labware accessories
    PCR cooling block

    CORNING: Corning® Thermowell ® GOLD 0.2 ml 8-Well PCR tube strips

    • Tube strips consist of eight 0.2 mL tubes connected together
    • Cap strips are sold separately
    • Made of thin wall polypropylene and designed for precise fit in heat blocks for optimum heat transfer
    • Certified DNase- and RNase-free
    • Autoclavable at 121°C

    Source: Website Merck KGaA

    Visit supplier website

    Arcis Sample Prep Kit 50rxn

    Arcis Sample Prep Kit is a ready to use kit comprising two reagent tubes enabling flexible processing of a variety of sample types and sample volumes including whole blood, saliva, bacteria, plant or animal cells. The fast and convenient Arcis DNA Sample Prep kit allows you to go from cells to downstream nucleic acid investigations without the need for isolation or purification.

    Source: Website Arcis Biotechnology

    Visit the website of the supplier
    Arcis Sample Prep Kit (50 rxn) consists of Lysis Buffer and Wash Buffer.

    Eppendorf, 500 µl Deepwell Plate

    Eppendorf® Deepwell plates, standard, 96 wells blue plate, conical bottom colorless wells, capacity 500 μL

    Source: Website Sigma-Aldrich

    Visit supplier website

    Download

    Download this App Note as PDF file

    Download PDFNucleic acid extraction with the Arcis Sample Prep Kit

    Customer’s voice

    Easy and flexible pipetting robot! The device is easy to use and therfore it saves us a lot of work. In addition, pipetting is more precise that manual pipetting and there is less risk of contamination. The software for making protocols is very inuitive and provides a lot of options, so the Assist can be easily used for different applications.

    Bianca Van Driel, Genmab