High Sensitivity D1000 ScreenTape Assay for TapeStation Systems

Quick Guide

The Agilent 4150 (G2992AA) and 4200 (G2991AA and G2991BA) TapeStation systems are automated platforms for scalable, flexible, fast, and reliable electrophoresis of nucleic acids. This Quick Guide is intended for use with the Agilent 4150 and 4200 TapeStation systems only. A Quick Guide specific for use with the Agilent 2200 TapeStation system is available online. The High Sensitivity D1000 Screen Tape assay is designed for analyzing double-stranded DNA molecules from 35 to 1000 basepairs.

Specifications

Analytical specifications

Analytical specifications	High Sensitivity D1000 ScreenTape assay
Sizing range	35 – 1000 bp
Typical resolution	35–300 bp: 15%, 300 – 1000 bp: 10%
Sensitivity¹	5 pg/µL
Sizing precision²	5% CV
Sizing accuracy²,³	±10%
Quantitative precision	15% CV
Quantitative accuracy²	±20%
Quantitative range	10 – 1000 pg/µL
Maximum buffer concentration in sample	7 mM KCl, 20 mM phosphate buffer, 20 mM guanidine-HCl, 80 mM NaCl, 20 mM NaOAc

Physical specifications

Physical specifications
Analysis time	16 samples: <20 min, 96 samples: <105 min
Samples per consumable	16
Sample volume required	2 µL
Kit stability	6 months
Kit size	112 samples

¹ Signal-to-noise >3 (single peak)

² Measured using one ladder per ScreenTape device

³ Sizing Accuracy for analysis with electronic ladder: ±20%

Storage Conditions

Reagent vials and ScreenTape devices: 2 – 8 °C (36 – 46 °F).
Store partially used ScreenTape devices upright at 2 – 8 °C (36 – 46 °F) for a maximum of 2 weeks.
Never freeze ScreenTape devices. Discard any accidentally frozen ScreenTape devices.

Kit Components

Part Number	Name	Color	Amount
5067-5584	High Sensitivity D1000 ScreenTape		7 ScreenTape devices
5067-5585	High Sensitivity D1000 Reagents	●	2 vials
	High Sensitivity D1000 Ladder	●	20 µL
5067-5603	High Sensitivity D1000 Sample Buffer	●	1 vial, 300 µL
5067-5587	High Sensitivity D1000 Ladder	●	1 vial, 20 µL

Limited Use Label License

Some products within this assay contain SYBR™ Green I, which is licensed from Life Technologies Corporation for use in research and development only. SYBR™ is a registered trademark of Life Technologies Corporation.

For Research Use Only
Not for use in Diagnostic Procedures.

Additional Material Required for Analysis with the TapeStation Systems

Loading tips (5067-5598, 1 pk or 5067-5599, 10 pk)
Optical Tube 8x Strip (401428) and Optical Tube Cap 8x Strip (401425)
Vortex mixer IKA MS3 with 96-well sample plate adapter
96-well sample plates (5042-8502) and 96-well Plate Foil Seal (5067-5154) (4200 TapeStation systems only)

Additional Equipment Required (Not Supplied)

Volumetric micropipettes for handling volumes from 1 to 15 µL
Centrifuges for tube strips and 96-well sample plates

WARNING Toxic agents

Refer to product material safety datasheets for further information.

When working with the ScreenTape assay follow the appropriate safety procedures such as wearing safety goggles, laboratory gloves and protective clothing.

CAUTION Damage to the TapeStation systems

Only use the recommended consumables and reagents with the TapeStation systems.

Essential Measurement Practices

Read about good measurement practices in the Agilent Information Center and/or in the System Manual.


Environmental conditions	Ambient operating temperature: 17 – 37 °C (63 – 99 °F) Keep reagents during sample preparation at room temperature
Steps before sample preparation	Allow Sample Buffer to equilibrate at room temperature for 30 min prior to use Vortex each vial and briefly spin down Flick Screen Tape device to eliminate bubbles in the buffer chamber
Pipetting practice	Pipette reagents carefully against the side of the 96-well sample plate or sample tube Ensure that no sample or Sample Buffer remains within or on the outside of the tip Care must be taken due to viscosity of the Sample Buffer
Mixing and centrifugation recommendations	Apply foil seal to 96-well sample plate or cap the tube strips prior to mixing and centrifugation Centrifuge to collect liquid at the base, then vortex using the IKA MS3 vortexer and adaptor at 2000 rpm for 1 min Briefly centrifuge and visually confirm that all liquid is collected at the bottom of the 96-well sample plate or tube strips and any air bubble is removed Run samples immediately after preparation

Ladder Considerations

Ladder is exclusively loaded from location A1 on the tube strip holder.
Always use a complete tube strip when running ladder or samples from the tube strip holder.
For best sizing and molarity quantification results, a ladder per ScreenTape device is recommended. Alternatively, an electronic ladder is available, which can be selected in the Agilent TapeStation Controller software.

High Sensitivity D1000 ScreenTape Assay Operating Procedure

Allow High Sensitivity D1000 Reagents to equilibrate at room temperature for 30 minutes.
Launch the Agilent TapeStation Controller software.
Flick the High Sensitivity D1000 ScreenTape device and insert it into the ScreenTape nest of the TapeStation instrument.
Select required sample positions in the TapeStation Controller software.
The required consumables (tips, further ScreenTape devices) are displayed in the TapeStation Controller software.
Vortex reagents and samples. Spin down before use.
Prepare ladder:
- For 2 ScreenTape devices: pipette 4 µL High Sensitivity D1000 Sample Buffer ● and 2 µL High Sensitivity D1000 Ladder ● at position A1 in a tube strip.
- For 1 ScreenTape device: pipette 2 µL High Sensitivity D1000 Sample Buffer ● and 4 µL High Sensitivity D1000 Ladder ● at position A1 in a tube strip.
- For more than 2 ScreenTape devices¹: pipette 15 µL High Sensitivity D1000 Sample Buffer ● and 15 µL High Sensitivity D1000 Ladder ● at position A1 in a tube strip.
For each sample, pipette 2 µL High Sensitivity D1000 Sample Buffer ● and 2 µL DNA sample in a tube strip or 96-well sample plate¹.
Apply caps to tube strips and/or foil seals to 96-well sample plates.
Mix liquids using the IKA MS3 vortexer at 2000 rpm for 1 min.
Spin down samples and ladder for 1 min.

Sample Analysis

Load samples into the TapeStation instrument. Place ladder in position A1 on tube strip holder.
Carefully remove caps of tube strips. Visually confirm that liquid is positioned at the bottom.
Click Start.
The TapeStation Analysis software opens automatically after the run and displays results.

Diagram Description: The diagram illustrates the sample preparation steps: Pipetting Sample Buffer (2 µL) and DNA (2 µL) into a tube, flicking the ScreenTape device, vortexing for 1 minute, spinning down for 1 minute, and finally placing samples, tips, and the ScreenTape device into the instrument.

Technical Support and Further Information

For technical support, please visit www.agilent.com/chem/contactus. Visit Agilent Technologies' web site. It offers useful information, support and current developments about the products and technology: www.agilent.com/genomics/tapestation.

¹ Agilent 4200 TapeStation system only

www.agilent.com

Agilent Technologies Inc. 2018-2021
Printed in Germany, Edition: 01/2021

Part No: G2991-90131 Rev. C
Document No: SD-UF0000091 Rev. C

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