Page 1
o
r useµPlate G0.5
E
N)ns for use
Eppendorf® μPlate G0.5
Instructions for use
Page 2
Copyright© 2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced
without the prior permission of the copyright owner.
Trademarks
Eppendorf
Hamburg, Germany.
KimWipe
Excel
Trademarks are not marked in all cases with ™ or
®
and the Eppendorf Logo are registered trademarks of Eppendorf AG,
®
is a registered trademark of Kimberly-Clark Corporation, Irving, TX, USA.
®
is a registered trademark of Microsoft Corporation, Redmond, WA, USA.
®
in this manual.
6144 900.209-01/062013
Page 3
Table of contents
Eppendorf
®
µPlate G0.5
English (EN)
Table of contents
1 Operating instructions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.1 Danger symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.2 Danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.1 Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.2 Delivery package . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3 Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.1 Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.2 User profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.3 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.3.1 Personal injury. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.3.2 Damage to device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.1 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.2 System requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3
5 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1 Applications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.1 Nucleic acid quantification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.2 Preparing measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.2.1 Defining the blank value procedure . . . . . . . . . . . . . . . . . . . . . . . 10
5.2.2 Average blanking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.2.3 Individual Blanking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2.4 Sample ID function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.2.5 Show Raw Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.2.6 Editing sample types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
5.2.7 Loading samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.2.8 Loading samples using the pipetting aid . . . . . . . . . . . . . . . . . . . 17
5.2.9 Pipetting with multi-channel pipettes . . . . . . . . . . . . . . . . . . . . . . 17
5.2.10 Pipetting with single-channel pipettes . . . . . . . . . . . . . . . . . . . . . 17
5.3 Performing a measurement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
5.4 Quality control of the µPlate G0.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
5.4.1 Average blanking out of range (CV ≥ 10 %) . . . . . . . . . . . . . . . . . 21
5.4.2 Ending the blanking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Page 4
Table of content s
Eppendorf
4
®
µPlate G0.5
English (EN)
6 Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
6.1 Working with the µPlate G0.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
7 Maintenance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
7.1 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
7.2 Disinfection/Decontamination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
8 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
8.1 Weight/dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
8.2 Additional parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
9 Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
9.1 Calculating the nucleic acid concentration . . . . . . . . . . . . . . . . . . . . . . . . . . 26
9.1.1 Example . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
9.2 Blank values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
9.2.1 Determining the average blank value . . . . . . . . . . . . . . . . . . . . . . 27
9.2.2 Determining the individual blank values. . . . . . . . . . . . . . . . . . . . 27
9.3 Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
9.3.1 Calculations based on an average blank value . . . . . . . . . . . . . . . 27
9.3.2 Calculations based on an individual blank value. . . . . . . . . . . . . . 28
10 Ordering Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
11 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
11.1 Shipping the µPlate G0.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Page 5
Operating instructions
Eppendorf
®
µPlate G0.5
English (EN)
1 Operating instructions
1.1 Using this manual
Read this operating manual completely before using the device for the first time.
Please also note the operating instructions for the accessories, if applicable.
This operating manual is part of the product. Thus, it must always be easily accessible.
Enclose this operating manual when transferring the device to third parties.
If this manual is lost, please request another one. For the current version, please refer
to our webpage www.eppendorf.com/worldwide
www.eppendorfna.com
(North America).
1.2 Danger symbols and danger levels
The safety instructions in this manual appear with the following danger symbols and
danger levels:
1.2.1 Danger symbols
Biohazard Explosion
Electric shock Toxic substances
Hazard point Material damage
(international) or
5
1.2.2 Danger levels
DANGER Will lead to severe injuries or death.
WAR NIN G Can lead to severe injuries or death.
CAUTION Can lead to light to moderate injuries.
ATTENTION May lead to material damage./Paragraph
1.3 Symbols used
Depiction Meaning
1.
2.
Actions in the specified order
Actions without a specified order
• List:
Text Display or software texts
Additional information
Page 6
Product description
Eppendorf
6
®
µPlate G0.5
English (EN)
2 Product description
2.1 Introduction
The µPlate G0.5 is a microvolume plate for quantifying small nucleic acid volumes (2 µL)
in the absorbance mode of PlateReader AF2200.
In the µPlate G0.5, 16 different samples can be analyzed at the same time during one
measuring procedure. After the measurement, which is controlled by the PlateReader
AF2200 software, the nucleic acid content is calculated automatically and the purity is
checked using the 260/280 ratio. The results will be output in an Excel spreadsheet. A
blank measurement, as well as a reference wavelength ratio at 340 nm at the start of the
measuring procedure, acts as a quality control check for the entire plate and displays any
pipetting or cleaning errors which may be present. The plate was specially developed for
the unique requirements of research laboratories, where various types of small sample
volumes are quantified.
2.2 Delivery package
Quantity Description
1µPlate G0.5
1 Pipetting template
1 Instructions for use
1Storage case
The µPlate G0.5 is compatible with the Eppendorf PlateReader AF2200.
The µPlate G0.5 is not included in the Eppendorf PlateReader AF2200 delivery package.
Page 7
Safety
Eppendorf
®
µPlate G0.5
English (EN)
3Safety
3.1 Intended use
The µPlate G0.5 is designed for use in molecular biology, biochemistry and cell biology
research laboratories. The µPlate G0.5 is exclusively intended for use indoors. The µPlate
G0.5 is designed as an auxiliary aid of the PlateReader AF2200 for laboratory
measurements for quantifying small nucleic acid volumes (2 L) in the absorbance mode.
3.2 User profile
The device and accessories may only be operated by trained and skilled personnel.
Before using the device, read the operating manual carefully and familiarize yourself with
the device's mode of operation.
3.3 Warnings for intended use
3.3.1 Personal injury
WARNING! Damage to health from toxic, radioactive or aggressive chemicals
as well as infectious liquids and pathogenic germs.
Observe the national regulations for handling these substances, the biological
security level of your laboratory, the material safety data sheets and the
manufacturer's application notes.
Wear personal protective equipment.
For comprehensive regulations about handling germs or biological materials
of the risk group II or higher, please refer to the "Laboratory Biosafety
Manual" (source: World Health Organisation, Laboratory Biosafety Manual, in
its current version).
7
WARNING! Damage to health due to contaminated device and accessories.
Decontaminate the device and the accessories before storage and shipping.
Page 8
Safety
Eppendorf
8
English (EN)
®
µPlate G0.5
3.3.2 Damage to device
NOTICE! Damage from the use of aggressive chemicals.
Do not use any aggressive chemicals on the device or its accessories, such as
strong and weak bases, strong acids, acetone, formaldehyde, halogenated
hydrocarbons or phenol.
If the device has been contaminated by aggressive chemicals, clean it
immediately using a mild cleaning agent.
NOTICE! Corrosion due to aggressive cleaning agents and disinfectants.
Do not use corrosive cleaning agents, aggressive solvents or abrasive
polishes.
Do not incubate the accessories in aggressive cleaning agents or disinfectants
for longer periods.
Page 9
Installation
Eppendorf
®
µPlate G0.5
English (EN)
4 Installation
4.1 Preparing installation
Store the transport packaging and packing material for future safe transport or
storage.
Use the details included in the delivery package to check that the delivery is complete .
Check all parts for any transport damage.
4.2 System requirements
The following items are required for a measurement with the µPlate G0.5:
• An Eppendorf PlateReader AF2200
• A computer with the PlateReader AF2200 software
Make sure that the following absorbance filters are installed in the filter slide of your
device:
• Position 1: 260 (5) nm
• Position 2: 280 (5) nm
• Position 3: 340 (10) nm
The PlateReader AF2200 and µPlate G0.5 may only be used at room temperature
under normal laboratory conditions.
9
Page 10
10
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5Operation
5.1 Applications
5.1.1 Nucleic acid quantification
For the quantification procedure in the µPlate G0.5, a sample volume of 2 L is sufficient
in order to achieve accurate results. The absorbance of the nucleic acid samples is
measured at 260 nm. The optical path length of the µPlate G0.5 is 0.5 mm. To assess the
purity of the nucleic acid, an additional measurement is conducted at 280 nm to display
any proteins that may be present in the sample. A 260/280 ratio between 1.8 and 1.9 is
acceptable for pure nucleic acids. A ratio < 1.8 may show that the sample contains
proteins or other impurities. If this is the case, additional purification steps are
recommended.
5.2 Preparing measurements
5.2.1 Defining the blank value procedure
The user can select from two options: determining the average blank value ("average
blanking") and determining the individual blank values ("individual blanking", set as
default).
5.2.2 Average blanking
To determine the average blank value, select the wells to be used for blanking. To do so,
drag a border around the corresponding sample positions in the plate preview. We
generally recommend carrying out the blank measurement with all 16 sample positions.
However, at least two wells are required to calculate an average value which will then be
used for the blank value correction of all measured samples.
To ensure reliable measuring results, the OD results measured must have a CV of < 10%
(CV= Coefficient of variation).
Abb. 5-1:"Start Blanking" button
Fig. 5-1: "Start Blanking" button
Page 11
Operation
Eppendorf
®
µPlate G0.5
English (EN)
(see Fig. 5-12 on p. 21)The CV value displayed in the "Last Blanking" field (see
Figure 13) is the coefficient of variation of all wells that were used for blanking.
5.2.3 Individual Blanking
All wells which will be used for the subsequent measurements will be included in the
determination of individual blank values (individual blanking). For each well used, the
information of the corresponding blank value is saved. The blank value correction of the
samples is carried out using the corresponding individual blank value (of the affected
well) instead of an average blank value.
Abb. 5-2:Individual Blanking checkbox
11
Fig. 5-2: Individual Blanking checkbox
We recommend using "Individual Blanking" as the default option for blanking because
this mode delivers the most precise and reliable results.
The blanking information is retained in the software until:
• The PlateReader is disconnected from the computer
• The software is exited.
• A different sample type is selected
If other sample types are selected, the blanking procedure must be repeated with the
corresponding wavelengths.
Page 12
12
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.2.4 Sample ID function
The user can use the "Sample ID" function to assign a unique name to each sample and
well. When the "Sample ID" button is clicked, a window will open where the user can
enter the desired sample names. Clicking on the "Clear" button will delete all entered
sample IDs.
Click on "Save" to save the entries and click on "Close" to end the procedure. Sample IDs
can also be copied from Excel worksheets and added to the sample ID list.
Abb. 5-3:"Sample ID" list
Fig. 5-3: "Sample ID" list
5.2.5 Show Raw Data
The µPlate G0.5 tab in the "Data Presentation" dialog box (in the Settings menu) contains
the "Show Raw Data" checkbox which can be used to output all measuring results as raw
data.
Page 13
Abb. 5-4:"Show Raw Data" checkbox
Eppendorf
Operation
®
µPlate G0.5
English (EN)
13
Fig. 5-4: "Show Raw Data" checkbox
If this box is selected, the worksheet with the results summary also contains the
absorbance raw data for all measured wavelengths, the absorbance values which were
corrected using the blank value and reference wavelength, the sample concentrations that
were calculated automatically and the ratios (260/280 ratio).
Page 14
14
Operation
Eppendorf
®
µPlate G0.5
English (EN)
Abb. 5-5:Raw data output in the Excel results summary
Fig. 5-5: Raw data output in the Excel results summary
Page 15
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.2.6 Editing sample types
In the "Sample Type" dropdown menu, select the "Others” option. The “Edit Samples”
window opens. The asterisk marks an open row in which additional sample types can be
entered with the corresponding standard coefficients [g/mL] for 1 cm layer thickness at
260 nm.
For example, a ratio wavelength of 280 nm can be selected from the selection menu.
You can delete samples by clicking on the row and pressing "Delete".
Abb. 5-6:Selecting and adding samples
15
Fig. 5-6: Selecting and adding samples
The correct filters must always be selected and they must be correctly defined in
the filter carriage. If an unavailable wavelength is selected (because the filter is
not in the slide), an error message appears.
Page 16
16
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.2.7 Loading samples
1. Open the µPlate G0.5.
2. Pipette the samples into the wells.
Abb. 5-7:µPlate G0.5 without pipetting aid
Fig. 5-7: µPlate G0.5 without pipetting aid
3. Close the µPlate G0.5 lid immediately after loading.
4. Insert the µPlate G0.5 in the PlateReader AF2200.
The filled µPlate G0.5 should always be measured within 5 minutes in order to
prevent evaporation of the sample or the blank - and avoid imprecise results.
Use the pipetting aid if you need assistance with precise application.
Page 17
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.2.8 Loading samples using the pipetting aid
1. Position the pipetting aid on the wells with the wells facing downward.
2. Secure the pipetting aid using the pins.
3. Pipette the samples into the wells.
Abb. 5-8:µPlate G0.5 with pipetting aid
Fig. 5-8: µPlate G0.5 with pipetting aid
4. Carefully remove the pipetting aid using an upward motion without touching the
samples.
5. Close the µPlate G0.5 lid immediately after loading.
6. Insert the µPlate G0.5 in the PlateReader AF2200.
The filled µPlate G0.5 should always be measured within 5 minutes in order to
prevent evaporation of the sample or the blank - and avoid imprecise results.
17
5.2.9 Pipetting with multi-channel pipettes
The quickest way to apply the 16 samples on the µPlate G0.5 is using an 8-channel
pipette.
5.2.10 Pipetting with single-channel pipettes
Loading with a single-channel pipette is also possible. In the process, the following points
must be observed:
• Always use a new, unused tip to prevent contamination with other samples.
• Work efficiently or the samples may evaporate. This will lead to inaccurate results.
• Close the µPlate G0.5 lid immediately after applying the samples.
Page 18
18
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.3 Performing a measurement
1. Start the PlateReader software.
2. Connect the software to the PlateReader.
The PlateReader software standard window opens.
3. In the Explorer bar, select "UV 260 nm microvolume" on the left-hand side of the
window by double clicking or via drag and drop.
Abb. 5-9:Method strip UV 260 nm microvolum e
Fig. 5-9: Method strip UV 260 nm microvolume
The corresponding method strip appears.
4. Select the desired mode for the blanking. If individual blank values are to be
determined, select the "Individual Blanking" checkbox. If you would like to determine
the average blank value, leave the checkbox unchecked (see Average blanking on p. 10)
and (see Individual Blanking on p. 11).
5. Select a sample type (e.g., dsDNA, ssDNA, RNA, etc.) from the drop-down list.
6. Click on the "Start Blanking" button to initialize the blank measurement.
The plate transport will move out of the instrument. The user will be prompted to insert
the µPlate G0.5 with the corresponding blank solution.
The blank measurement starts and can be monitored in the window with the progress bar.
The results of the blanking (date and time, sample positions selected for the blanking,
blank value range and maximum CV) are displayed next to the plate preview in the
method strip and stored until the device is disconnected.
If the blank measurement has been completed successfully, the plate will move out
automatically. Now the plate is ready for samples to be deposited and analyzed.
Remove the remaining blank solution from the sample positions by wiping off the quartz
wells using a piece of lint-free paper. Then place 2 L of each sample on each of the wells.
If the µPlate G0.5 has been loaded with samples and correctly positioned on the plate
carrier, click on the green "Start" button.
Page 19
Operation
Eppendorf
®
µPlate G0.5
English (EN)
An Excel worksheet opens automatically in the background while the measurement is
running. All measuring results (including the automatically calculated nucleic acid
concentration and the 260/280 ratio) will be shown condensed in a table (similar to the
plate layout). The absorbance values for each sample will also be shown with all relevant
wavelengths.
Abb. 5-10:Excel result sheet
19
Fig. 5-10: Excel result sheet
The plate will be automatically moved out as soon as the measuring procedure has been
completed. A message appears asking the user if he/she would like to perform another
measurement.
If additional (identical) measurements are to be conducted, wipe off all sample residue
from the previous measurement completely and place new samples on the plate. Click on
"Yes" to start the measurement.
If no additional measurements will be conducted, click on "No". A special worksheet,
containing a summary of the results of all previous measurements, appears in the Excel
workbook.
Page 20
Operation
Eppendorf
20
English (EN)
Abb. 5-11:Automatic summary of measuring results
®
µPlate G0.5
Fig. 5-11: Automatic summary of measuring results
If all measurements have been completed, thoroughly clean the µPlate G0.5(see Cleaning
on p. 24). Please store the µPlate G0.5 properly.
Excel workbooks that contain measuring results are not saved automatically.
This must be done by the user.
Page 21
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.4 Quality control of the µPlate G0.5
5.4.1 Average blanking out of range (CV ≥ 10 %)
If the coefficient of variation of the average blanking is out of range, the failed wells are
highlighted in pink.
The wells highlighted in pink are contaminated with lint, finger prints, etc.
In addition, an error message appears requesting the user to repeat the blanking
measurement.
Abb. 5-12:Pink color code
21
Fig. 5-12: Pink color code
Page 22
22
Operation
Eppendorf
®
µPlate G0.5
English (EN)
5.4.2 Ending the blanking
There are two ways to end the blanking:
1. Repeat the blanking using the same plate but with white wells only (e.g., D1 – E2, as
shown in the figure above). Drag a frame around these wells.
Abb. 5-13:Purple color code
Fig. 5-13: Purple color code
The newly selected wells appear white; the deviating wells switch from pink to violet.
All other wells remain blue, which indicates that they are not being used.
2. Move the plate out, repeat the cleaning procedure, load the plate with fresh blanking
solution and start the blanking over again.
Page 23
Troubleshooting
Eppendorf
®
µPlate G0.5
English (EN)
6 Troubleshooting
6.1 Working with the µPlate G0.5
Always switch on the PlateReader AF2200 before loading the µPlate G0.5 with samples.
Work efficiently when loading the µPlate G0.5 with samples to prevent the samples from
evaporating.
Only use the µPlate G0.5 at room temperature. Otherwise, significant fluctuations in
temperature may change the optical path length and, therefore, affect the absorbance
values.
23
Page 24
Maintenance
Eppendorf
24
®
µPlate G0.5
English (EN)
7 Maintenance
7.1 Cleaning
Cleaning the µPlate G0.5 is particularly important in order to achieve optimal measuring
results. The following two procedures are used to clean the µPlate G0.5.
7.1.1 Cleaning in the ultrasonic bath
1. Fill an ultrasonic bath with water and place a suitable beaker, filled with distilled water,
inside it.
2. Switch on the ultrasonic, immerse the lid of the µPlate G0.5 in the beaker and move the
lid back and forth in the beaker for about 20 seconds. Make sure that the plate hinge is
not immersed in the process.
3. Repeat this procedure with the lower part of the µPlate G0.5.
4. Remove excess water from the µPlate G0.5 using dry and oil-free compressed air.
7.1.2 Cleaning with Kimwipe cleaning cloth
1. Moisten a Kimwipe lab cleaning cloth with 70 % ethanol and use it to clean the inner
and outer surfaces of the µPlate G0.5.
2. Moisten a piece of cotton or a Kimwipe cleaning cloth with distilled water and use it to
clean all quartz wells on both sides of the µPlate G0.5.
3. Use a dry Kimwipe cleaning cloth to wipe off excess liquid.
Store the plate at a dust and lint-free location after cleaning. The quartz wells must be free
of lint, dirt and streaks. Contamination may lead to incorrect measurements. If many
different samples are measured one after the other, the quartz wells can be cleaned using
a (moist) Kimwipe cleaning cloth. The correct cleaning and care is important to extend
the service life of the µPlate G0.5 and reduce the amount of maintenance required. We
recommended to store the cleaned µPlate G0.5 in the storage case.
Lint, dirt or fingerprints on the quartz wells can significantly alter the OD values.
Prevent dirt from entering the spacer bars because this could lead to a change to
the high-precision spacer on the lid of the µPlate G0.5, which would affect the
OD values.
Samples may only be loaded on clean quartz wells.
7.2 Disinfection/Decontamination
All parts of the µPlate G0.5 that come into contact with patient samples, positive
control samples or dangerous materials must be treated as potentially infectious
areas.
1. Spray or apply 70% ethanol, which is typically used for laboratory cleaning
procedures, on the entire plate.
2. After an exposure time of 5 minutes, dry the µPlate G0.5 using a lint-free Kimwipe
cleaning cloth.
Page 25
Technical data
Eppendorf
®
µPlate G0.5
English (EN)
8 Technical data
8.1 Weight/dimensions
WeightμPlate G0.5
Weight approx. 160 g
DimensionsμPlate G0.5
Length 127.8 mm
Width 85.4 mm
Height 14.6 mm
8.2 Additional parameters
Physical
Optics 16 quartz wells (one per sample)
Quartz lens Optical path length: 0.5 mm
Diameter: 2.2 mm
Parallel measurement 16 sample positions (2 rows of 8 samples
each)
Sample volume Min. 2 µL
25
Wavelength settings
Eppendorf PlateReader AF2200 260 nm (5) nm
280 nm (5) nm
340 nm (10) nm
Measuring time for the entire plate
Nucleic acid quantification 1:15 min for all 16 samples
Typical performance data
Wavelength absorbance
Absorbance bandwidth Depends on filters used
Detection limit (DNA concentration) 1 ng/µL dsDNA
Reproducibility of a sample (50 µg/mL) < 1 % CV
260/280 ratio (50 µg/mL) ± 0,07
Precision at 260 nm < 0.2 %
Accuracy at 260 nm < 0.5 %
Page 26
Calculations
Eppendorf
26
®
µPlate G0.5
English (EN)
9 Calculations
With each measurement, an additional measurement is performed automatically at a
reference wavelength of 340 nm to correct any absorbance value errors which may have
been caused by dirt on the outer surfaces of the Quartz wells.
9.1 Calculating the nucleic acid concentration
After the absorbance measurements of the nucleic acids have been performed in the
µPlate G0.5, the PlateReader AF2200 software and Excel automatically calculate the
nucleic acid concentration according to the Beer–Lambert law, including the reference
values.
A = ε * d * c
A Absorbance
ε
Molar extinction coefficient (L mol
-1
cm-1)
D Distance (path length in cm)
c
Concentration (mol L
-1
)
Generally, the absorbance A is defined in analytical chemistry as:
A
= log10(I0/I) [OD],
λ
whereby I is the intensity of transmitted light of a specified wavelength which passed
through a sample and I
is the intensity of the light before it entered the sample.
0
Absorbance measurements are frequently used in analytical chemistry because the
absorbance of a sample changes depending on the thickness of the sample and the
concentration of the absorbing species in the sample. Absorbance is a logarithmic
dimension and the unit is [A].
9.1.1 Example
An absorbance value of 1 OD equals a transmission of 10 %; an absorbance value of 2 OD
equals a transmission of 1 %, etc. As absorbance calculations are based on logarithmic
dimensions, calculations between absorbance sample values and absorbance blank values
are done by division instead of subtraction. Additional information can be found in the
literature on the Beer–Lambert law.
Page 27
Calculations
Eppendorf
®
µPlate G0.5
English (EN)
9.2 Blank values
9.2.1 Determining the average blank value
The average absorbance value at 340 nm, which was determined for all wells used for
blanking, is subtracted from the average absorbance value at 260 nm or 280 nm. The
relative deviation of the wells used to determine the average blank value must be under 10
% to ensure that a measurement can be started.
Ext
average blank value
= Ext
260 average
– Ext
340 average
[OD]
9.2.2 Determining the individual blank values
The well-specific absorbance value at 340 nm is subtracted from the corresponding
absorbance value at 260 nm.
Ext
blank value A1
Ext
blank value A2
Ext
blank value B1
= Ext
= Ext
= Ext
260 A1
260 A2
260 B1
– Ext
– Ext
– Ext
340 A1
340 A2
340 B1
[OD]
[OD]
[OD]
etc.
For each well used, the corresponding blank value is saved. The blank value correction for
each sample is carried out using the corresponding individual blank value (of the affected
well) instead of an average blank value. Each well that is used for the sample
measurement must have been previously taken into account for blank value
determination.
27
9.3 Samples
9.3.1 Calculations based on an average blank value
The well-specific absorbance value at 340 nm is subtracted from the corresponding
absorbance value at 260 nm. Then, a value is determined using the average value for each
well used for the sample measurement.
Ext
Ext
Ext
= (Ext
A1
= (Ext
A2
= (Ext
B1
260 A1
260 A2
260 B1
– Ext
– Ext
– Ext
340 A1
340 A2
340 B1
) – Ext
average blank value
) – Ext
) – Ext
average blank value
average blank value
[OD]
[OD]
[OD]
etc.
Page 28
Calculations
Eppendorf
28
®
µPlate G0.5
English (EN)
9.3.2 Calculations based on an individual blank value
The well-specific absorbance value at 340 nm is subtracted from the corresponding
absorbance value at 260 nm. Then, a value is determined using the corresponding value
for each well used for the sample measurement.
Ext
Ext
Ext
A1
A2
B1
= (Ext
= (Ext
= (Ext
260 A1
260 A2
260 B1
– Ext
– Ext
– Ext
340 A1
340 A2
340 B1
) – Ext
A1 individual blank value
) – Ext
A2 individual blank value
) – Ext
B1 individual blank value
[OD]
[OD]
[OD]
etc.
The absorbance values at 280 nm will also be corrected using the corresponding
absorbance values at 340 nm. The corrected absorbance values will be used to calculate
the 260/280 ratio.
Page 29
Eppendorf
10 Ordering Information
Order no.
(International)
Order no. (North
America)
Description
μPlate G0.5
Eppendorf microvolume plate for the
6144 000.003 6144000003
Eppendorf PlateReader AF2200
PlateReader AF2200
6141 000.002 230 V / 50 – 60 Hz
6141 000.010 6141000010 120 V / 50 – 60 Hz
μPlate G0.5 and PlateReader AF2200
(bundle)
Eppendorf microvolume plate and
PlateReader AF2200
6141 000.908 230 V / 50 – 60 Hz
6141 000.909 120 V / 50 – 60 Hz
6141000922 120 V / 50 – 60 Hz
UV/Vis filter slide for PlateReader
AF2200
Preconfigured filter slide, optimized
for applications in the UV and Vis
6141 070.019 6141070019
range, 4 filters (260, 280, 340, 600 nm)
Filterslide case for PlateReader
6141 070.035 6141070035
AF2200
Ordering Information
®
µPlate G0.5
English (EN)
29
Page 30
30
Transport
Eppendorf
®
µPlate G0.5
English (EN)
11 Transport
11.1 Shipping the µPlate G0.5
If you intend to ship the µPlate G0.5 to Eppendorf Service, observe the following items:
• A contaminated µPlate G0.5 poses a health risk.
• Follow the instructions in the decontamination certificate that is available as a PDF on
the Eppendorf webpage: www.eppendorf.com/decontamination.
• Decontaminate the µPlate G0.5.
• Include the entire decontamination certificate for returned goods with the shipment,
including the serial number of the µPlate G0.5.
Page 31
Page 32
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