Automated reagent and sample aliquoting with the ASSIST PLUS pipetting robot

  • Automated reagent and sample aliquoting with the ASSIST PLUS pipetting robot

    Aliquoting samples made easy

    Aliquoting is a routine pipetting procedure that refers to the transfer of one or multiple portion(s) of a liquid to a different vessel. It is a useful technique to create space-saving back-up plates of primary tubes, or to split expensive reagents into smaller volumes to increase overall open vial stability. When done manually, aliquoting can be time consuming and error prone. Combining the ASSIST PLUS pipetting robot with the VOYAGER adjustable tip spacing pipette or D-ONE single channel pipetting module provides unique solutions for the user. With the VOYAGER, the pipetting robot offers adjustable tip spacing when aliquoting between different types of labware, while the D-ONE pipetting module guarantees precise aliquoting from a single tube.

  • Table of contents

    Aliquoting is a routine pipetting procedure that refers to the transfer of one or multiple portion(s) of a liquid to a different vessel. It is a useful technique to create space-saving back-up plates of primary tubes, or to split expensive reagents into smaller volumes to increase overall open vial stability. When done manually, aliquoting can be time consuming and error prone. Combining the ASSIST PLUS pipetting robot with the VOYAGER adjustable tip spacing pipette or D-ONE single channel pipetting module provides unique solutions for the user. With the VOYAGER, the pipetting robot offers adjustable tip spacing when aliquoting between different types of labware, while the D-ONE pipetting module guarantees precise aliquoting from a single tube.

Key benefits

  • The ASSIST PLUS pipetting robot ensures accurate sample transfers to help prevent human errors.
  • Using the VOYAGER adjustable tip spacing pipette to aliquot samples between different labware formats in high throughput makes the process simple and easy.
  • Reagent aliquoting from tubes provided in a kit saves money and time when using the D-ONE single channel pipetting module.
  • The operator gains more walk-away time and is protected from exposure to harmful substances by using the ASSIST PLUS pipetting robot.

Downloads: App note and protocols for the automated reagent and sample aliquoting with the ASSIST PLUS pipetting robot

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.

Overview: How to aliquot reagents and samples faster

In this application note, we show six examples of reagent and sample aliquoting into different labware types (Figure 1 shows single and multiple aliquoting):

  1. Sample (from swab tubes) aliquoting for space-saving, long-term storage
  2. Aliquoting of compounds dissolved in DMSO for cryogenic storage
  3. Purified nucleic acid aliquoting from deep well extraction plate
  4. Sample (bronchial lavage, urine, etc.) splitting for biobanking 
  5. Reagent (primers, probes, etc.) splitting to increase overall open vial stability
  6. Aliquoting reagents (buffers, master mixes, etc.) for kit preparation

 

Experimental set-up

Schematic of single and multiple aliquoting of samples between different labware types.
Figure 1: Illustration of a single aliquoting versus a multiple aliquoting procedure.

Step-by-step procedure:

 

1. Sample (from swab tubes) aliquoting for space-saving, long-term storage

Aliquoting from swab tubes to a 96 deep well plate.

Place an INTEGRA rack for swab tubes with patient samples on deck position B (Figure 2, blue) and a 96 deep well plate on deck position C (Figure 2, magenta).

Run the VIALAB program 'Aliquoting_from_swab_tubes'. The 6 channel 1250 µl VOYAGER adjustable tip spacing pipette with 1250 µl Sterile, Filter, Wide Bore GRIPTIPS® aspirates 1000 µl of each sample from the swab tubes (Figure 3a) and dispenses them to the 96 deep well plate (Figure 3b), automatically changing the tips between different samples. After filling up the first half of the 96 deep well plate, the pipette prompts the user to exchange the INTEGRA rack for swab tubes on deck position B (Figure 2, blue). After loading the next 48 swab tubes, the VOYAGER continues to fill up the second half of the 96 deep well plate.

Tips:

  • A 50 µl air gap at the end of every aspiration step minimizes the risk of cross-contamination.
  • Wide bore tips and slower aspiration/dispense speeds can be used when samples tend to be more viscous.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with INTEGRA rack for swab tube, a 96 deep well plate and the VOYAGER pipette.
Figure 2: Deck set-up for sample transfer from swab tubes to 96 deep well plate. Position A: Empty. Position B: INTEGRA rack for swab tubes (blue). Position C: 96 deep well plate (magenta).
A close-up view of the ASSIST PLUS pipetting robot transferring samples from swab tubes to 96 deep well plate.
Figure 3: Sample transfer from a) swab tubes to b) 96 deep well plate with VOYAGER pipette.

2. Aliquoting of compounds dissolved in DMSO for cryogenic storage

Aliquoting from a flat bottom 96 well plate to cryogenic vials.

Place a flat bottom 96 well plate with samples on deck position B (Figure 4, blue) and an INTEGRA rack for cryogenic tubes on deck position C (Figure 4, magenta).

Run the VIALAB program 'Aliquoting_from_f-bottom_plate'. The 8 channel 300 µl VOYAGER pipette with 300 µl Sterile, Filter, Low Retention GRIPTIPS aspirates 200 µl of each sample from the flat bottom 96 well plate and dispenses them to 1.5 ml cryogenic vials. Tips are changed automatically between samples. The pipette will inform the user to change the INTEGRA rack for cryogenic vials on deck position C (Figure 4, magenta) when the first half of the 96 well plate has been aliquoted.

Tips:

  • Setting an aspiration/dispense delay ensures that viscous liquid enter and exit the GRIPTIPS properly.
  • Low Retention GRIPTIPS and slower dispense speeds can be used to ensure the full volume of liquid is expelled.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with a flat bottom 96 well plate, an INTEGRA rack with cryogenic tubes, and the VOYAGER pipette.
Figure 4: Set-up for sample transfer from flat bottom 96 well plate. Position A: Empty. Position B: flat-bottom 96 well plate (blue). Position C: INTEGRA rack for cryogenic tubes (magenta).

3. Purified nucleic acid aliquoting from deep well extraction plates

Aliquoting from deep well extraction plate to 1.5 ml microcentrifuge tubes.

Place a deep well extraction plate on deck position B (Figure 5, blue) and an INTEGRA rack for 1.5 ml microcentrifuge tubes on deck position C (Figure 5, magenta).

Run the VIALAB program 'Aliquoting_from_extraction_DWP'. The 8 channel 125 µl VOYAGER pipette with 125 µl Sterile, Filter GRIPTIPS aspirates 50 µl of each sample from the elution plate and dispenses them into 1.5 ml microcentrifuge tubes. Tips are changed automatically between samples. After half of the samples have been transferred, the pipette will inform the user to load a new INTEGRA rack with empty 1.5 ml microcentrifuge tubes on deck position C (Figure 6, magenta) and continue aliquoting.

Tip:

  • The procedure parameters can be easily modified in the VIALAB software. If the volume of the purified nucleic acid is different, this can be adjusted rapidly.

 

Graphical representation of the ASSIST PLUS pipetting robot deck set-up with a deep well extraction plate, an INTEGRA rack with microcentrifuge tubes, and the VOYAGER pipette.
Figure 5: Deck set-up for purified nucleic acid transfer from deep well extraction plates. Position A: Empty. Position B: Deep well extraction plate (blue). Position C: INTEGRA rack for 1.5 ml microcentrifuge tubes (magenta).

4. Sample (bronchial lavage, urine, etc.) splitting for biobanking

Multiple sample aliquoting from centrifuge tubes to FluidX™ tubes.

Place an INTEGRA rack for centrifuge tubes with patient samples on deck position A (Figure 6, blue), and a FluidX 96-format tubes on positions B and C (Figure 6, magenta).

Run the VIALAB program 'Aliquoting_from_centrifuge_tube'. The 6 channel 1250 µl VOYAGER pipette with 1250 µl Sterile, Filter GRIPTIPS aspirates the samples from the centrifuge tubes, and dispenses 250 µl of each sample to FluidX tubes until every sample contains eight 250 µl aliquots, with automatic changing of tips between different samples.

Tip:

  • The repeat dispense function of an electronic pipette speeds up this process, making it four times faster than using a mechanical multichannel pipette.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with an INTEGRA rack with centrifuge tubes, a 96-format FluidX tubes plate, and the VOYAGER pipette.
Figure 6: Deck set-up for sample aliquoting from centrifuge tubes to FluidX™ tubes. Position A: INTEGRA rack for centrifuge tubes. Position B: FluidX 96-format tubes (magenta). Position C: FluidX 96-format tubes (magenta).

5. Reagent (primers, probes, etc.) splitting to increase overall open vial stability

Multiple aliquoting from a 2 ml manufacturer's tube to 0.5 ml microcentrifuge tubes.

Place an INTEGRA rack for 2 ml tubes with the manufacturer's tube on position A1 on deck position B (Figure 7, blue) and an INTEGRA rack for 0.5 ml centrifuge tubes on deck position C (Figure 7, magenta).

Run the VIALAB program 'Aliquoting_from_manufacturer_tube'. The 5-1250 µl D-ONE pipetting module with 1250 µl Sterile, Filter GRIPTIPS aspirates the reagent from the manufacturer’s tube and dispenses 50 µl into every 0.5 ml microcentrifuge tube (Figure 8).

Tip:

  • This protocol is optimized for speed. If the reagent needs to be aliquoted at high precision, the tip type can be switched to the 125 µl GRIPTIPS in VIALAB, and the system will automatically adjust the volume that needs to be aspirated.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with two INTEGRA racks with microcentrifuge tubes, and the D-ONE pipetting module.
Figure 7: Set-up for reagent transfer from manufacturers’ tubes to microcentrifuge tubes. Position A: Empty. Position B: INTEGRA rack for 2 ml tubes. Position C: INTEGRA rack for 0.5 ml microcentrifuge tubes.
A close-up view of the ASSIST PLUS pipetting robot aliquoting reagent from a manufacturer’s tube with the D-ONE pipetting module.
Figure 8: Reagent aliquoting from a manufacturer tube with the D-ONE pipetting module.

6. Aliquoting reagents (buffers, master mixes, etc.) for kit preparation

Multiple aliquoting from 100 ml INTEGRA reservoir to 2 ml tubes.

Place a 100 ml INTEGRA reservoir on deck position A (Figure 9, blue), and INTEGRA racks for 2 ml tubes with screw caps on deck positions B and C (Figure 9, magenta).

Run the VIALAB program 'Aliquoting_from_reservoir'. The 8 channel 1250 µl VOYAGER pipette with 1250 µl Sterile, Filter GRIPTIPS aspirates the reagent from the reservoir, and dispenses 1000 µl into each 2 ml tube.

Tip:

  • If the aliquoting volume needs to be smaller than 500 µl, the dispense type can be switched in VIALAB, speeding up the process.
Graphical representation of the ASSIST PLUS pipetting robot deck set-up with an INTEGRA reservoir, two INTEGRA racks with microcentrifuge tubes, and the VOYAGER pipette.
Figure 9: Deck set-up for reagent aliquoting from an INTEGRA reservoir to tubes. Position A: INTEGRA 100 ml reservoir. Position B: INTEGRA rack for 2 ml centrifuge tubes (magenta). Position C: INTEGRA rack for 2 ml centrifuge tubes (magenta).

Remarks

  • VIALAB software: The VIALAB software gives the option to change protocols and labware according to workflow needs, adapting easily to future requirements.
  • Partial plates: Programs can be adapted to accommodate a different number of samples at any time.

Conclusion

  • The ASSIST PLUS pipetting robot equipped with the VOYAGER adjustable tip spacing pipette is the perfect solution to transfer samples and reagents between different labware formats using single or multi-dispensing.
  • The D-ONE pipetting module enables fast and easy multiple aliquoting from a single manufacturer’s tube to any other labware type.
  • The automated protocols provided offer simple, fast, and ready-to-use solutions for six common aliquoting procedures with maximum walk-away time.
  • Thanks to its compact footprint, the ASSIST PLUS can be placed in a laminar flow hood whenever sterile conditions are required during aliquoting.

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ASSIST PLUS, Pipetting Robot D-ONE Single Channel Pipetting Module, 0.5-300 µl VOYAGER, 8 Channel, 125 µl VOYAGER, 8 Channel, 300 µl VOYAGER, 6 Channel, 1250 µl VOYAGER, 8 Channel, 1250 µl Tip deck for D-ONE on ASSIST PLUS Rack for 1.5/2 ml microcentrifuge tubes Rack for 0.5 ml microcentrifuge tubes Rack for 15 ml centrifuge tubes, 4x6 tubes Rack for cryogenic tubes, 6x8 tubes Rack for swab tubes GRIPTIP, 125 µl, Sterile, Filter GRIPTIPS®, 300 µl, Sterile, Filter GRIPTIPS®, 1250 µl, Sterile GRIPTIPS®, 1250 µl, Sterile, Filter, Wide Bore Multichannel Reagent Reservoir, 100 ml, PS Multichannel Reagent Reservoir, 100 ml Greiner Bio-One: Masterblock, 96 well, 2 ml, V-bottom Greiner Bio-One: CRYO.S, 2 ml Greiner Bio-One: 96 Well Microplate, PS, F-Bottom Greiner Bio-One: Reaction Tube, 1.5 ml, PP, Natural, Attached Cap Greiner Bio-One: Reaction Tube, 0.5 ml COPAN: Swab tubes VWR: Centrifuge tubes, Sterile, PP 0.7ml Dual-coded Tube, 96-format

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,
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  • error free pipetting.

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ASSIST PLUS Pipetting Robot

D-ONE Single Channel Pipetting Module, 0.5-300 µl

The D-ONE single channel pipetting module offers hands-free transfers from individual tubes or wells on the ASSIST PLUS pipetting robot. This makes it the perfect add-on for labs that want to go beyond the possibilities of multichannel pipetting to access unlimited applications. It is designed to offer easy automation of normalization, hit picking and master mix preparation, freeing up your time while eliminating transcription errors.

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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.

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

VOYAGER, 8 Channel, 300 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. 4723

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VOYAGER, 8 channel series

VOYAGER, 6 Channel, 1250 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.

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VOYAGER, 6 channels, 1250μL

VOYAGER, 8 Channel, 1250 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.

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Tip deck for D-ONE on ASSIST PLUS

  • Supports accommodation of 1 x 12.5 µl/ 125 µl tip box and 1 x 300 µl/ 1250 µl tip box
  • Compatible with D-ONE Pipetting Module 0.5 - 300 µl and 5 - 1250 µl
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Rack for 1.5/2 ml microcentrifuge tubes

Hold up to 48 tubes on removable carriers for easy loading and storage.

Part No. 4540

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Rack for 0.5 ml microcentrifuge tubes

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Microcentrifuge tube rack fitting 0.5 ml, 1.5 ml and 2 ml tubes.

Rack for 15 ml centrifuge tubes, 4x6 tubes

This tube rack is designed to hold 15 ml centrifuge tubes in a 4x6 ANSI/SLAS format.

Part No. 4542

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Rack for 1.5/2 ml microcentrifuge tubes

Rack for cryogenic tubes, 6x8 tubes

Part No. 4544

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Rack for cryogenic tubes, 6x8 tubes

Rack for swab tubes

Part No. 4546

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Rack for swab tubes (13x75 mm), 6x8 tubes

GRIPTIP, 125 µl, Sterile, Filter

GRIPTIPS® 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.

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GRIPTIPS®, 300 µl, Sterile, Filter

GRIPTIPS® 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.

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GRIPTIPS®, 1250 µl, Sterile

GRIPTIPS® 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.

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1250 µl, Non-sterile, GripTips

GRIPTIPS®, 1250 µl, Sterile, Filter, Wide Bore

GRIPTIPS® 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.

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Multichannel Reagent Reservoir, 100 ml, PS

  • 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
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Part No. 4320, 4322

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Reagent reservoirs 10ml, 25ml, 100ml

Multichannel Reagent Reservoir, 100 ml

Users benefit by reusing the sturdy base and tossing the disposable insert. Reservoirs 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. Unlike traditional reservoirs that have hard to see graduations molded directly into the plastic, INTEGRA reservoirs are made of crystal clear polystyrene and fit into a reusable base with bold, crisp, clearly visible graduation markings. The unique design leads to more accurate reagent measurements, no over pouring, and less waste.

  • Dead volume of only 310 µl
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Greiner Bio-One: Masterblock, 96 well, 2 ml, V-bottom

96 Well Polypropylene MASTERBLOCK®, natural

  • Free of detectable DNase, RNase, human DNA
  • Non-pyrogenic 
  • Manufactured under DIN ISO 9001 guidelines
  • Can be traced all the way back to production through a defined LOT number system
  • Ideal microplates for storing sample material but are also ideal for cultivating bacteria or yeasts
  • Compatible with automated systems
  • Uniform external dimensions and tolerances
  • Alphanumeric well coding
  • High chemical resistance
  • High temperature resistance  (-196°C to + 121°C)
  • Sealable with adhesive films and heat sealer
  • Solid bottom
  • Well profile: V-bottom (pyramidal shape)
  • Volume: 2 ml

Part No. 780270

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Greiner Bio-One: CRYO.S, 2 ml

Cryo.s™ Freezing Tubes, 2 ml 

  • Free of detectable DNase, RNase, human DNA
  • Non-Pyrogenic, non-cytotoxic
  • CE-marked
  • Made of high-grade polypropylene which guarantees high thermal resistance in a temperature range from -196°C to +121°C
  • As a saftety precaution Cryo.s™ should be stored in a freezer or exclusively in the gas phase over the liquid nitrogen
  • Suitable for shipment by air
  • Round bottom
  • Working volume: up to 2.2 ml
  • Diameter: 12.5 mm; total height incl. lid: 47 mm
  • With starfoot base and external thread
  • With red screw cap
  • With white, scratch-resistant writing area
  • Sterile 
  • With 150 inserts per case (for additional labelling on the cap)
  • Additional inserts Cat.-No. 304 171 (50 pieces per bag)

Part No. 126280

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Greiner Bio-One: 96 Well Microplate, PS, F-Bottom

  • Free of detectable DNase, RNase, human DNA
  • Non-pyrogenic
  • Manufactured under DIN ISO 9001 guidelines
  • Can be traced all the way back to production through a defined LOT number system
  • Is ideal for precise optical measurements.
  • The measuring light source is not deflected by the well profile
  • Excellent optical properties as a result of the flat bottom of the wells
  • For microscopic apllications (bottom reading)
  • Solid bottom Well profile: F-bottom/standard (flat bottom)
  • Sterile

Part No. 655161

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Greiner Bio-One: Reaction Tube, 1.5 ml, PP, Natural, Attached Cap

  • Free of detectable DNase, RNase, human DNA
  • Non-Pyrogenic
  • High chemical and temperature resistance
  • Volume: 1.5 ml
  • With graduation and attached cap
  • Suitable for Eppendorf

Part No. 616201

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Reaction tube, 1,5 ml, PP, natural

Greiner Bio-One: Reaction Tube, 0.5 ml

  • Free of detectable DNase, RNase, human DNA
  • Non-Pyrogenic
  • High chemical and temperature resistance
  • Volume: 0.5 ml
  • With attached cap
  • Suitable for Viatron

Part No. 667201

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COPAN: Swab tubes

ESwab™ combines a COPAN-invented flocked swab with 1mL of Liquid Amies in a plastic, screw cap tube. The innovative system elutes over 90% of patient specimen into the liquid medium. The result is evidence-based, improved pathogen recovery, expanded testing capabilities, and better patient care.

Part No. 490CE.A

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VWR: Centrifuge tubes, Sterile, PP

Centrifuge tubes, Ultra-High Performance, VWR®

These leakproof, conical bottom centrifuge tubes prevent breakage and leakage during high speed centrifugation, and are made of non cytotoxic, medical grade resin that allows users to easily see sample volume and colour.

Part No. 525-1084

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0.7ml Dual-coded Tube, 96-format

Azenta Life Sciences dual-coded tubes feature a 2D code and human readable number on tube base, allowing compatibility with low throughput manual, semi-automated or fully automated workflows on integrated platforms

  • Next Generation manufacturing techniques
  • Code readable without removing tubes from racks
  • Suitable for cryogenic storage
  • Made from high-quality virgin PP
  • Available in 96-well format SBS racks or bulk
  • Sealable with TPE/auto friendly screw caps

Part No. 66-62318-Y6

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0.7ml Dual-coded Tube, 96-format, Internal Thread

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The most versatile and affordable liquid handling system on the market!

"The ASSIST PLUS coupled with the D-ONE pipette makes my Pharmacogenomic normalization workflow a piece of cake! "

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