tecan NanoQuant Plate Quick Manual

NanoQuant Plate™

Quick Guide

2 Quick Guide NanoQuant PlateTM No.30035094 Rev No. 1.4 2013-06

Table of Contents

1. General Information ...................................................................... 3

1.1 Introduction ....................................................................... 3

1.2 Contents of the NanoQuant Plate Package.................... 3

1.3 Computer Requirements .................................................. 4

1.4 System Requirements ...................................................... 5

1.5 Applications ...................................................................... 6

2. Measurement Procedure............................................................... 7

2.1 Software Installation Procedure ...................................... 7

2.2 Performing a Measurement ............................................. 7

2.3 File menu ......................................................................... 15

2.4 Quality Control of NanoQuant Plate ............................. 17

3. NanoQuant Plate .......................................................................... 19

3.1 Parameters ...................................................................... 19

3.2 Handling and Cleaning ................................................... 21

3.3 Applying Samples ........................................................... 22

3.4 NanoQuant Plate Disinfection ....................................... 23

4. Calculations ................................................................................. 25

4.1 Calculation of Nucleic Acid Concentration .................. 25

5. About the Quick Guide ................................................................ 29

2013-06 Quick Guide NanoQuant PlateTM No.30035094 Rev No. 1.4 3

1. General Information

1.1 Introduction

Tecan’s NanoQuant Plate is intended as a general laboratory measurement
tool for the quantification of small volumes (2μl) of nucleic acids in absorbance
mode and the measurement of the labeling efficiency of nucleic acids labeled
with fluorescent dyes. The NanoQuant Plate permits the application and
parallel measurement of 16 different samples in a single measurement
procedure. After the measurement, which is controlled by Tecan’s i-control
software, the calculation of nucleic acid content and purity check using the
260/280 ratio is performed automatically and the results are displayed in an
Excel sheet. A blanking measurement, including an integrated reference
wavelength at the beginning of the measurement procedure, functions
simultaneously as a quality control check for the entire plate and indicates any
pipetting or cleaning mistakes. The plate has been designed to meet the
requirements of research laboratories working with various types of low-volume
samples including fluorophore-labeled nucleotides.

1.2 Contents of the NanoQuant Plate Package

The NanoQuant Plate package for Infinite reader series contains the following
items:

• NanoQuant Plate

• Pipetting Aid

• Safety Certificate

• This Quick Guide

• Storage Box

The NanoQuant Plate is available for the following readers:

• Infinite M200 PRO

• Infinite F200 PRO

• Infinite M200 PRO NanoQuant

• Infinite F200 PRO NanoQuant

• Infinite M1000

• Infinite F500

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1.3 Computer Requirements

The following computer requirements are needed to use the i-control software:

Minimum Recommended

PC

Windows XP/Vista (32 bit)/Windows 7 (32 or 64 bit)
Windows compatible PC with a Pentium compatible
processor running at 1 GHz

2 GHz
(Dual Core)

Operating
System

Windows XP (32-bit) SP3
Windows Vista (32-bit)
Windows 7 (32-bit)
Windows 7 (64-bit)

Windows XP
(32-bit) SP3

Memory

Windows XP: 512 MB RAM
Windows Vista (32-bit): 1 GB RAM
Windows 7 (32-bit): 1 GB RAM
Windows 7 (64-bit): 2 GB RAM

1 GB RAM
2 GB RAM
2 GB RAM
3 GB RAM

Space
Requirements

700 MB 1 GB

Monitor Super VGA Graphics

Resolution 1024 x 768 1280 x 1024

Color Depth 256

Mouse Microsoft mouse or compatible pointing device

Communication 1 x USB 2.0

2 x USB 2.0,
1 x RS232
(Serial)

Devices

1 x CD-ROM drive
Windows Vista: DirectX 9 graphics and 32 MB of

graphics memory (for Home Basic); 128 MB of

graphics memory plus WDDM support for all other
versions

Windows 7: DirectX 9 graphics device with WDDM 1.0
or higher driver

.NET

Microsoft .NET Framework 2.0
If this version is not present, the install/upgrade
program will install it side-by-side with any existing
installations of the .NET Framework.

Windows
Installer

3.1
If this version is not present, the install/upgrade

program will install it.

Microsoft Excel

2002, 2003, 2007, 2010 (32-bit) – Starter edition NOT
supported!

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1.4 System Requirements

To perform a NanoQuant measurement, the following items are required:

Infinite 200 Pro

• An Infinite 200 Pro reader with firmware version V2.0 or higher

• A computer with i-control V1.4 Service Pack 1 or higher installed

• The NanoQuant Plate Package

Make sure the following absorbance filters are available on the filter slide of
your Infinite F200 PRO:

• Position 1: 260 nm (5 nm bandwidth)

• Position 2: 280 nm (3 nm bandwidth)

• Position 3: 340 nm (10 nm bandwidth)

• Position 4: free (for individual use)

For Infinite F200 PRO, the filter positions must remain in the order in which they
were delivered. The original filter positions guarantee the fastest filter switching
for well-wise measurements. The Infinite M200 PRO can be used immediately
for measurement without any calibration of the monochromator.

Infinite M1000, Infinite F500

• An Infinite M1000 reader (REF 30061442) with main firmware version
V2.0 or higher

• An Infinite F500

• Software: i-control V1.8 or higher

• The NanoQuant Plate Package

Make sure the following absorbance filters are available on the filter slide of
your Infinite F500:

• Position 1: 260 nm (5 nm bandwidth)

• Position 2: 280 nm (3 nm bandwidth)

• Position 3: 340 nm (10 nm bandwidth)

Note

Only use the Infinite reader and the NanoQuant Plate at room temperature and
under normal laboratory conditions.

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1.5 Applications

Nucleic Acid Quantification (Infinite 200 PRO, Infinite M1000,
Infinite F500)

For the quantification procedure in the NanoQuant Plate, a sample volume of
2 μl is sufficient for accurate results. Absorbance of nucleic acid samples is
measured at 260 nm. The optical path length of the NanoQuant Plate is

0.5 mm. To assess the purity of the nucleic acid, an additional measurement at

280 nm is performed to indicate proteins in the sample. For pure nucleic acids,
a 260/280 ratio between 1.8 - 1.9 is acceptable. If this ratio is lower than 1.8 it
may indicate the presence of proteins or other contaminants. If this is the case,
an additional purification step/procedure is recommended.

Labeling Efficiency (Infinite M200 PRO and Infinite M1000 only)

Working with nucleic acids labeled with fluorescent dyes requires samples of
high quality. Besides common nucleic acid quantification and nucleic acid purity
check with 260/280, the labeling efficiency is an important criterion for improved
research results. With the NanoQuant Plate it is possible to measure
absorbance of nucleic acids labeled with Cy3, Cy5, Alexa 555, Alexa 647 and
many other fluorescent dyes.

Note

All measurements on Infinite M1000 involving NanoQuant Applications are
performed using one measurement channel only.

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2. Measurement Procedure

2.1 Software Installation Procedure

The i-control software is installed using the following procedure:

1. Insert the i-control software CD ROM into your CD ROM drive.

2. A window opens with different selectable options.

3. Choose Software and install i-control.

4. Follow the instructions of the Wise Installation Wizard.

5. When installation is successful, exit the Installation window.

6. Switch on and connect to Infinite instrument.

Note

i-control is delivered with the Infinite reader series.

2.2 Performing a Measurement

For applications using the NanoQuant Plate, a tab called Applications is
implemented in i-control software, so that all measurements can be
performed quickly and easily.

1. Start i-control.

2. Connect to the Infinite instrument. The standard i-control window
opens.

3. Select Applications in the lower left part of the window.

Figure 1: Overview of i-control script

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4. Select the desired measurement type in the control bar on the left side
of the window by double-clicking or dragging and dropping:

• Nucleic acid quantification (Infinite 200 PRO, Infinite M1000,

Infinite F500)

• Labeling Efficiency (Infinite M200 PRO and Infinite M1000 only)

5. The corresponding measurement stripe appears and the NanoQuant

Plate definition file (NanoQuant Plate Tecan 16 Flat Black) is

automatically selected in the Plate field.

6. Select blanking mode. Select the Individual Blanking check box for
individual blanking or leave the check box clear for average blanking.
See Individual Blanking and Average Blanking on page 12.

7. Depending on the connected instrument, the wavelengths used for
measurement are selected automatically (make sure that the correct
filters are properly installed and defined on the filter slides of the
Infinite F200 PRO and Infinite F500 instrument).

Instrument Bandwidth at 260 nm Bandwidth at 280 nm

Infinite M200 PRO: 5 nm 5 nm

Infinite F200 PRO: 5 nm 3 nm

Infinite M1000: 5 nm 5 nm

Infinite F500: 5 nm 3 nm

8. Select a sample type (e.g. dsDNA, ssDNA, RNA, etc. in the Sample
type drop-down list.

9. In addition, select the respective dye(s) in the Markers drop-down lists
for Labeling Efficiency measurements. If the samples are labeled with
only one fluorophore, set the drop-down list of Dye 2 to None.

10. When all settings are correct, click the Start Blanking button to
initialize the blanking measurement. The plate transport moves out
and the user is requested to insert the NanoQuant Plate with the
respective blanking buffer.

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Figure 2: Correct orientation of NanoQuant Plate in the reader

11. The first measurement step is blanking with the required buffer. A
separate window opens and the blanking procedure can be observed.

12. The blanking measurement is started and can be monitored in the
measurement progress window. If blanking has been performed
successfully, the sample positions are highlighted in yellow and the
screen color changes to a homogenous green (Nucleic Acid

Quantification) or blue (Labeling Efficiency). Blanking results (date and

time, samples positions that were selected for blanking, blanking
range, and maximum CV) are displayed next to the plate preview in
the measurement stripe and saved until the instrument is
disconnected.

13. When the blanking measurement has been completed successfully,
the plate is moved out automatically. The plate is now ready for
sample application and analysis. The green Start button is now
accessible.

14. Remove any remaining blanking buffer from the sample positions by
wiping the quartz spots with a piece of lint free paper and apply 2 μl of
the samples onto each spot.

15. When the NQP is loaded with samples and correctly placed onto the
plate carrier, click the green Start button.

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16. As the measurement is performed an Excel sheet opens automatically
in the background. All measurement results (including the
automatically calculated nucleic acid concentration, the 260/280 ratio,
and for Labeling Efficiency measurements, the dye concentration) are

concisely displayed in a matrix (analogous to the plate layout). The

OD values of each sample at all relevant wavelengths are also
displayed.

Figure 3: Overview of Excel result sheet

17. Once the measurement procedure is finished, the plate is moved out
automatically. A pop-up message appears, asking if the user wishes
to perform another measurement.

• If additional (identical) measurements are to be performed, wipe

o any sample residues from the previous measurement and

apply new samples. Click

Yes to start the measurement.

• If no further measurement is to be performed, click No. An extra

sheet appears in the Excel workbook summarizing the results of
all previous measurements.