Bio-Rad SmartSpec Plus Spectrophotometer User Manual
SmartSpec
™
Plus
Spectrophotometer
Instruction Manual
Catalog Number
170-2525
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)
Table of Contents
Page
Section 1 General Safety Information .....................................................................1
1.1 Caution Symbol......................................................................................................1
1.2 Precautions.............................................................................................................1
Section 2 Packing List ..............................................................................................1
Section 3 Installation ................................................................................................2
3.1 Environmental Requirements .................................................................................2
3.2 Power On and Initial Display .................................................................................2
Section 4 Introduction ..............................................................................................3
4.1 Identification of System Components.....................................................................3
4.2 Setup ......................................................................................................................7
4.3 Serial Port Functions ............................................................................................10
4.4 PC Serial Connection Instructions........................................................................11
4.5 Key Pad Descriptions ...........................................................................................13
Section 5 Quick Guide............................................................................................15
Section 6 Operating Instructions ...........................................................................17
Section 7 Examples of Assays.................................................................................25
7.1 DNA/RNA ...........................................................................................................25
7.2 Protein Assay .......................................................................................................31
7.3 Spectral Scanning.................................................................................................44
7.4 Kinetics ................................................................................................................46
7.5 OD600..................................................................................................................47
7.6 Lambda (λ) ..........................................................................................................48
Section 8 Troubleshooting ......................................................................................49
Section 9 Maintenance............................................................................................50
9.1 Cleaning Up a Fluid Spill .....................................................................................50
9.2 Cleaning Quartz Cuvettes.....................................................................................51
9.3 Changing Thermal Printer Paper ..........................................................................51
Section 10 Specifications ..........................................................................................52
Section 11 System, Accessories, and Reagents for SmartSpec
Spectrophotometer .................................................................................53
11.1 Product Information .............................................................................................53
11.2 Bio-Rad Protein Assays........................................................................................53
11.3 Cuvette Selection Guide .......................................................................................54
11.4 Selecting the Appropriate Cuvette........................................................................54
For technical service call your local Bio-Rad office. In the U.S., call 1-800-4BIORAD
(1-800-424-6723).
Copyright© (2002) Bio-Rad Laboratories, Inc. All rights reserved.
Warranty
The SmartSpec Plus Spectrophotometer and accessories are warranted against defects
in materials and workmanship for one year. If any defects occur in the instruments or
accessories during this warranty period, Bio-Rad Laboratories will repair or replace the
defective parts at its discretion without charge. The following defects, however, are specifically excluded:
1. Defects caused by improper operation.
2. Repair or modification done by anyone other than Bio-Rad Laboratories or an autho-
rized agent.
3. Damage caused by substituting alternative parts.
4. Use of fittings or spare parts supplied by anyone other than Bio-Rad Laboratories.
5. Damage caused by accident or misuse.
6. Damage caused by disaster.
7. Corrosion caused by improper solvent†or sample.
For any inquiry or request for repair service, contact Bio-Rad Laboratories. Inform
Bio-Rad of the model and serial number of your instrument.
Important: This Bio-Rad instrument is designed and certified to meet EN61010
*
safety standards. Certified products are safe to use when operated in accordance with
the instrument manual. This instrument should not be modified or altered in any way.
Alteration of this instrument will:
Void the manufacturer's warranty
Void the EN61010 safety certification
Create a potential safety hazard
Bio-Rad Laboratories is not responsible for any injury or damage caused by the use of
this instrument for purposes other than those for which it is intended, or by modifications
of the instrument not performed by Bio-Rad Laboratories or an authorized agent.
†
SmartSpec Plus Spectrophotometer is not compatible with chlorinated hydrocarbons (e.g., chloroform), aromatic hydrocarbons
(e.g., toluene, benzene) or acetone. Use of organic solvents voids all warranties.
*
EN61010 is an internationally accepted electrical safety standard for laboratory instruments.
!
!
Section 1
General Safety Information
This instrument is intended for laboratory use only.
This product conforms to the "Class A" standards for electromagnetic emissions
intended for laboratory equipment applications. It is possible that emissions from this product may interfere with some sensitive appliances when placed nearby or in the same circuit
as those applications. The user should be aware of this potential and take appropriate measures to avoid interference.
1.1 Caution Symbol
Read the manual before using the SmartSpec Plus Spectrophotometer. For technical
assistance, contact your local Bio-Rad office or, in the U.S., call technical services at
1-800-4BIORAD (1-800-424-6723).
Definition of Symbols
Caution (refer to accompanying documents)
1.2 Precautions
1. Do not use near flammable materials.
2. Always inspect the SmartSpec Plus for damaged components before use.
3. Always connect the system to the correct AC power source.
4. Always connect the correct instrument (printer or computer only) via the serial port
connector.
5. Do not pour liquid into the sample chamber. Thorough clean-up is needed after each
spill (Section 9.1).
6. Do not place objects on the SmartSpec Plus.
7. The SmartSpec Plus has no customer-serviceable components, do not open the unit.
Section 2
Packing List
The SmartSpec Plus Spectrophotometer (catalog number 170-2525) is shipped with the
following components. Upon receiving your instrument, please check that all items listed
below were shipped. If any items are missing or damaged, contact your local Bio-Rad
office.
Item Quantity
Instruction manual 1
Warranty card (please complete and return) 1
SmartSpec Plus Spectrophotometer 1
Power cable 1
Cuvette spring 1
Thermal paper roll 1
1
!
!
Section 3
Installation
3.1 Environmental Requirements
To ensure correct operation and stable performance over an extended period of time,
install the SmartSpec Plus in a location which meets the following conditions:
• Room temperature between 15 and 35°C. Not recommended for cold room use (4°C).
• Not exposed to direct sun light.
• Not subject to direct or continuous vibration.
• Not subject to intense magnetic or electromagnetic fields.
• Relative humidity between 0–95%.
• Area free from corrosive gases or other corrosive substances.
• Area with very little dust or other airborne particles.
• Allow a 10 cm minimum space around the instrument for proper air flow.
3.2 Power On and Initial Display
1. Power up SmartSpec. When you turn on SmartSpec Plus, it will go through a
self-diagnostic routine during which it tests the lamp, optical components, diode array,
powersupply and other components. If a problem is detected, an error message will be
displayed and you should call Bio-Rad Technical Support. If boot up is successful and no
errors are detected, the Home screens will be displayed.
alternating with
2
CHOOSE ANY
SMARTSPEC ASSAY
04:23:11 PM SmartSpec
Fri Apr 23, 1999
Section 4
Introduction
4.1 Identification of System Components
Fig. 1. View of the SmartSpec Plus Spectrophotometer.
Instrument
1. Keypad interface Change menu and select options by pressing these
buttons. Press a single button to perform action
described on the menu display.
2. Display 2 x 24 alphanumeric backlit LCD display. Displays
various menus and options available.
3. Sample compartment door Lift door to insert cuvette into the cuvette chamber.
4. Integrated thermal printer Built-in alphanumeric/graphic printer. Press print
button to record results immediately.
5. Printer paper compartment Remove door to install new roll of printer paper
door
Fig. 2. Rear view of the SmartSpec Plus Spectrophotometer.
6. Fan Guard Exhaust air leaves cabinet through this opening.
7. Universal Power Input Module Appropriate power cord is connected to the power
and ON/OFF Switch input module. Instrument power switch.
8. Serial Output Port RS-232 connection to serial printer or computer.
3
1
2
5
4
3
8
7
6
Fig. 3. View of the SmartSpec Plus sample compartment.
9. Cuvette chamber Insert cuvette into the cuvette chamber. Ensure that
the cuvette is placed in the proper orientation with
respect to the light path.
10. Cuvette storage compartment Storage space for cuvette.
11. Cuvette spring Aligns cuvette. Accommodates various cuvette
configurations.
12. Light path Light path axis.
Fig. 4. View of the SmartSpec Plus integrated thermal printer paper with door removed.
13. Thermal printer paper
14. Printer paper shaft
15. Paper removal instructions
4
10
11
12
14
13
15
9
5
UV/Visible Spectroscopy is based on the absorption of light as a function of wavelength.
All spectrophotometers have a light source that generates light of specific wavelengths. The
SmartSpec Plus uses a xenon flash lamp as its light source. Xenon flash lamps have several
advantages over the conventional light sources in that they reduce warm up time, do not heat
up the instrument and have a lamp life of 10 to 15 years. Additionally, xenon flash lamps emit
both UV and visible wavelengths, in contrast to deuterium lamps which emit only UV
wavelengths or tungsten lamps which primarily emit visible wavelenghts. In the SmartSpec
Plus, the xenon flash lamp pulses light only when measuring absorbance of a sample.
Fig. 5. A symbolic representation of the SmartSpec Plus optics module.
The path of light in the SmartSpec Plus is as follows:
Light from the xenon flash lamp
Lens
Sample
Lens
Slit
Concave mirror
Grating
Concave mirror
Photodiode array detector
The advantages of using a grating as a dispersion device are that:
1. They eliminate nonlinear dispersion.
2. Are not temperature sensitive.
Photodiode arrays have several advantages:
1. They acquire spectral data simultaneously without mechanically moving parts;
2. Have excellent wavelength reproducibility and,
3. A wide dynamic range.
It is also important to note here that in an array, several photodiode detectors are
arranged on a silicon crystal. The added benefit of an array is the ability to do side-by-side
readings, thus increasing speed.
Source
Lens
Sample Slit
Concave
Mirror
Grating
Concave
Mirror
Detector
The Lambert-Beer’s law identifies the relation between sample concentration and the
intensity of transmitted light. Absorbances and % transmittances are defined by the following equations:
% Transmittance (%T)
%T = It/I0* 100 = 10
-εcl
* 100
Absorbance or optical density (A or O.D.)
A = log 101/T = log 10I0/It= log1010
εcl
= εcl
Where I0= Intensity of incident light
It= Intensity of transmitted light
ε = Molar extinction coefficient (liter/mole/cm)
c = Sample concentration (mol/liter)
l = Optical pathlength
Fig. 6. Lambert-Beer's law.
A = log 1/T A = εcl
Fig. 7. Relationship between Transmittance and Concentration and Absorbance and
Concentration.
6
Sample concentration C
ll
Incident light l
o
Transmitted light lt= lo10
-εεc
l
4.2 Setup
When the SmartSpec Plus is not carrying out an assay, you can enter the Setup routine
by pressing Setup. To proceed through Setup without changing the current setting, press
Enter. Most of the current settings can be changed by pressing Select until you see your
desired choice; sometimes you have to use the number pad to change an entry. In either
case, press Enter after the selection is made. When you have gone all the way through
Setup, SmartSpec Plus will automatically return to Home screen. You can exit Setup at any
time by pressing Cancel. When you exit Setup, all changes will be saved.
In Setup you can specify the following:
1. Language
2. Pathlength
3. Background compensation for DNA/RNA measurements.
4. Default type of nucleic acid.
5. Default method to determine DNA and RNA oligonucleotide molar extinction
coefficient and molecular weight.
6. Default number of sample replicates.
7. New operator name.
8. Current operator.
9. Serial port export.
10. Current time.
11. Current date.
12. Time display mode.
13. Date display mode.
14. Performance validation
1. Language. SmartSpec Plus may display one of six different languages on its LCD
screen. After pressing the Setup key, you can choose from: English(Eng),
French(Fra), German(Deu), Italian(Ita), Spanish(Esp) or Japanese.
English is the default language and will be automatically selected if no choice is made.
If another language is selected, once you exit Setup all changes will be saved and the
instrument will display the selected language until changed in the Setup.
2. Pathlength. The pathlength describes the distance that the light travels as it passes
through the sample, and is the same as the width of the cuvette. By using a cuvette with
a shorter pathlength, you can measure the absorbance of more concentrated solutions.
The relationship between absorbance, path length and sample concentration is described
by Beer’s law where
A = εbc
A is the absorbance, ε the molar extinction coefficient in liter/mole/cm, b the pathlength in cm and c the concentration in moles/liter.
Pressing Select once will change the screen to
As you continue pressing Select, the choices change to 2 mm and then 1 mm.
7
Select Pathlength:
>=Help 10
mm
Select Pathlength:
>=Help 5
mm
3. Background compensation. When using the DNA/RNA assay, you can choose to
have SmartSpec Plus automatically subtract background absorbance readings collected
at 320 nm from the A260 and A280 measurements. If the background subtraction option
is invoked, then the A320 is subtracted from the A260 and A280 readings before these
measurements are displayed and before they are used in the calculation of nucleic acid
concentration. The default condition is to not subtract the A320 background. You can
toggle background subtraction on and off by pressing Select.
4. Default type of Nucleic Acid. SmartSpec Plus can convert absorbance reading of nucleic
acid solutions into concentration. The conversion factor is determined by the type of
nucleic acid being measured. You can select the default type of nucleic acid in Setup,
so that when you press DNA/RNA, this type of nucleic acid will be shown first on the
display. You can choose another type of nucleic acid at any time by pressing Select.
The choices are double-stranded DNA (dsDNA), single-stranded DNA (ssDNA),
RNA, DNA oligo and RNA oligo. The factory default choice is dsDNA.
5. Default method to determine oligo molar extinction coefficient and molecular
weight. SmartSpec Plus uses the molecular weight and molar extinction coefficients of
oligonucleotides to convert the absorbance readings into mass (µg/ml) and molar
(pmol/µl) concentrations. You can input the molecular weight and molar extinction
coefficient directly if you know them, otherwise SmartSpec Plus will use one of three
methods to estimate them for you: Length, Composition or Sequence. The simplest
method to estimate either the molecular weight or the molar extinction coefficient is by
the length of the oligonucleotide. This method assumes an equivalent distribution of all
four nucleotides among the oligo. More accurate estimates can be calculated if you
specify the composition (i.e., the number of A’s, C’s, G’s and T’s or U’s) of the oligo
and the best estimate of molar extinction coefficient is calculated with the sequence of
the oligonucleotide.
Press Select to change the default to Composition (Compstn) and press Select again to
set the default to Sequence.
6. Default number of sample replicates. SmartSpec Plus has the ability to accept samples
(and standards) in replicate groups and to calculate a mean and standard deviation for
each sample (or standard) replicate group. The factory default number of sample
replicates is 1, but you can change the number of replicates to any number between 1 and 9.
Use the keypad to choose the number of sample replicates and press Enter or Setup
when finished. During most assays you can change the number of sample replicates by
choosing Options from the Ready Screen.
8
Substract A320 Background
for DNA/RNA? Yes
>=Help
Select default type of
DNA/RNA: dsDNA
>=Help
Select default method of
find oligo conc: length
Enter default number of
sample replicates:
7. New Operator. When SmartSpec Plus prints a report, it includes the name of the
operator (if specified). You can add a new operators name to the list of operators in
Setup.
Press Select to toggle from No to Yes and the screen will change to
The keyboard will automatically switch to the alpha mode and you can use the keys to
enter a name with as many as eight characters. To enter an A in the first space, press the
2 key once. To enter a B, press the 2 key twice, and to enter a C, press it three times. To
move from the first space to the second space, press the right arrow key. When the last
letter of the name has been specified, press Enter. If you want to mix numerals with letters, you can toggle back and forth between the alpha mode and number mode by pressing Alpha. When the LED on the Alpha key is lit, then the keyboard is in alpha mode.
You can enter a space by pressing the right arrow key twice.
8. Current operator. Press Select until the desired name is displayed on the screen and
then press Enter. This name will be printed on the final report.
9. Serial Port or Printer Data Export. This feature allows the user to export data using
either the printer, serial port or both. The screen will prompt for the desired configuration.
Use the Select key to toggle between the data export options.
10. Current time. The time will be displayed in the present format.
If the displayed time is incorrect, press Select and then Enter.
• Use the number pad to enter the hour and press Enter, then enter the minutes and press
Enter again. After setting the time, press Select to change from AM to PM if you are
using the 12-hour format. Press Enter when complete.
11. Current date. The date will be displayed in the present format.
If the date is not correct, press Select to toggle from Yes to No and Enter.
• Use the number pad to input the new date. Press Enter to move to the next field. Press
Select to set the day of the week. Press Enter when you are finished.
9
Want to add the name of
a new operator? No
Enter new operator's
name: (8 char):
Current time: 1:12 PM
is this OK? Yes
Printer: On
Serial port: Off
Enter new time. Select
AM or PM: ____ : ____ AM
Current date: (MM/DD/YY)
11/25/98 Is this OK? Yes
Enter new date in format
MM/DD/YY: ___/___/YY
12. Time convention. You may choose to print time in 12-hour or 24-hour mode. Toggle
between the choices by pressing Select. and then press Enter.
Press Select for
13. Date convention. You can choose to print the date in either MM/DD/YY or DD-MMYY mode. Toggle between the choices by pressing Select and then press Enter.
Press Select for
14. Performance Validation. The Performance Validation function requires the purchase
of the SmartSpec Plus with Performance Validation Package (170-2520) or the separate
Performance Validation Package (170-2509). SmartSpec Plus will automatically perform
absorbance and wavelength tests and create a printable report.
Press Enter to access the Performance Validation program. Enter the four digit
Validation Cuvette number found on the Cuvette's certificate of analysis.
After entering absorbance information found on the Validation Cuvette's certificate of
analysis, the SmartSpec Plus will automatically perform wavelength and absorbance
accuracy tests. This process can take up to 5 minutes to complete. When finished, a
report may be printed.
15. Exit Setup. After going through the entire Setup menu, the display will return to the
Home screen. All changes will be saved. You can also exit Setup at any time by
pressing Cancel and all changes up to that point will be saved.
4.3 Serial Port Functions
The SmartSpec Plus has the ability to communicate with an external printer or
computer via the serial port located at the back of the instrument. The SmartSpec Plus
sends data to these peripherals as ASCII text. When connecting the SmartSpec Plus to an
external printer or computer, a "standard serial" cable should be used. The correct pin-out
for this cable is described in the table below. Check the printer or computer for the correct
serial port connector end (male or female).
DB9F Connector Signal and DB9M Connector
to SmartSpec Plus (male) Direction to serial device
2 TX - - > 3
3 < - - RX 2
5 Common 7
(other pins are open in SmartSpec Plus)
10
Select date display mode
MM/DD/YY
e.g. 11/25/98
Select date display mode
DD-MM-YY
e.g. 25-11-98
Select time display mode
24-hr
e.g. 13:12
Select time display mode
12-hr
e.g. 1:12
The serial port parameters are: 9600 baud rate, 8 data bits, no parity, 1 stop, and
XON/XOFF flow control. These parameters are pre-set in the unit. When using a serial printer
set the printer to the serial port parameters specified above. Please note that the data are sent
from the SmartSpec Plus to a computer or serial printer as hex characters. If your computer or
printer is not set to accept these characters please refer to the instrument’s instruction manual
for correct settings. (For detailed instructions see Section 4.4.)
4.4 PC Serial Connection Instructions
1. Connect RS232 9-pin serial cable from SmartSpec Plus to computer.
2. Using a terminal program (e.g. HyperTerminal), set the connection to 9600 baud, 8 bitsno-parity-1 stop bit (9600-8-N-1), no flow control.
3. In addition to the actual data transmission/reception parameters, other parameters in
HyperTerminal may have to be set. The correct settings may vary from computer to
computer, but these are settings currently used and offered as an example. It may not be
necessary to implement all parameters. Only the Connect Direct to Com Port is essential.
From the main HyperTerminal menu
File
Properties
Open the properties dialog box. Using the tabbed HyperTerminal properties box:
Connect To
Connect using Direct to Com [n] (n = serial port number). This must be set.
Settings
Terminal Keys
Del
Emulation ANSI
Tel net terminal ANSI
Terminal Setup
✓✓
Underline
✓✓
Blink
Backscroll buffer line 500
ASCII Setup
✓✓
Echo typed characters locally
✓✓
Wrap lines that exceed terminal width
11
4. When the SmartSpec Plus sends data, either when requested from Scan, or Kinetics, or
when the user activates the data echo toggle in Setup, the data streams to the terminal
window.
5. Capturing data in HyperTerminal and saving the data in a file:
In the main HyperTerminal window menu, first click Transfer and then click Capture Text.
Transfer
Capture Text . . .
[give file name and path]
e.g. c:\program files\accessories\hyperterminal\capture.txt
A dialog box with the default file name, file extension and file path will appear. Accept
the default, or type in a new path and/or file name.
Use the SmartSpec and send data from the SmartSpec to the computer.
When finished:
Transfer
Capture Text . . .
Stop (closes file)
6. Start Excel (or Wordpad, or MS Word, etc.)
If using Excel click
File
Open
Open the SmartSpec/HyperTerminal data file.
e.g. c:\program files\accessories\hyperterminal\scan.txt
make sure files of types: (e.g. *.txt) is set to correct extension
Select and Open desired file
Text Import Wizard:
Delimited
Colon (:) is used as a delimiter for Scan and Kinetics
General data type should be sufficient for expressing numerical data. The system
selects the appropriate data type.
12
4.5 Keypad Descriptions
Print SmartSpec Plus will print data on the current sample,
a user-selectable range of samples or all samples in the
current assay.
Abs SmartSpec Plus will automatically display the
absorbance reading of the current sample and the
wavelength being read. If more than one wavelength
is being read, press Abs to toggle among the different
wavelengths. If you are reading one of a replicate
group, you can choose to see the absorbance of the
current replicate or you can see the mean and standard
deviation of all members of the replicate group by first
pressing Abs.
Conc The Conc key is used in the DNA Oligo and RNA
Oligo assays to toggle the display of concentration
from units of µg/ml to pmoles/µl. This key has no
function for the Scan, Kinetics and λ assays.
A260:A280 SmartSpec Plus will calculate and display the ratio of
A260 to A280. This is a method of approximating the
purity of nucleic acid preparations; pure preparations of
DNA have ratios of about 1.8 and pure RNA preparations
have a ratio of about 2.0. If you are using background
compensation (in Setup), then the ratio is calculated
after the background reading of A320 is subtracted
from the A260 and A280.
Dilution Factor The dilution factor is used as a final multiplier in
calculation of concentration from absorbance data.
The default dilution factor is 1.0 and the dilution factor
is reset to 1.0 each time that one of the Assay buttons
is pressed. The adjusted concentration will be printed
on the final report only if the dilution factor is changed
after measuring each unknown sample.
Setup SmartSpec Plus will enter the setup routine where you
can specify the pathlength, defaults for certain options,
time and date convention and time and date. You exit
Setup either by proceeding all the way through the
routine, or by pressing Cancel at any time. When you
exit the Setup routine, all changes are saved.
Assay Buttons The six assay buttons are in the second row of function
keys on the face of the SmartSpec Plus. Press one of
these to begin an assay with SmartSpec Plus. When
any assay button is pushed, SmartSpec Plus resets the
sample number to 1, and checks the memory for
unprinted data.
DNA/RNA SmartSpec Plus will automatically collect absorbance
data at 260 and 280 nm, and use those readings to
calculate nucleic acid concentration. If selected in
Setup, background will automatically be subtracted at
320 nm. The calculation of concentration depends on
the type of nucleic acid specified.
13
Protein SmartSpec Plus will automatically read one or more
wavelengths depending on the type of assay specified
(Bradford, Lowry, BCA or UV Protein) or the user
can specify a wavelength. SmartSpec Plus will
convert absorbance data to concentration by use of a
standard curve, except for the case of UV Protein in
which the absorbance readings are multiplied by
user-selectable constants. The standard curve can be
one recalled from memory, or a new one constructed
by SmartSpec Plus.
Scan SmartSpec Plus will collect and print the absorbance
of a sample over a user-defined range of wavelengths
from 200 to 800 nm. An optional background
wavelength may be specified and its absorbance will
be subtracted from all absorbances in the scan range
before the scan is displayed. SmartSpec Plus will
identify and quantitate the points of maximum and
minimum absorbance over the scanned range. These
data can be printed or downloaded via the serial port
to an external computer.
Kinetics SmartSpec Plus will collect data from a single cuvette
at fixed periodic intervals over a certain time period.
Both the interval of data collection and length of data
collection are specified by the user. An optional
background wavelength may be specified. The kinetic
readings may be printed or exported to external
software for analysis.
OD600 SmartSpec Plus will collect absorbance data at 600
nm. This assay is used for monitoring the growth of
bacterial cultures. The user may specify a conversion
factor which is multiplied by the dilution factor and
the absorbance to calculate the concentration in cells
per ml.
λλ
SmartSpec Plus will collect data at up to three different
user-specified wavelengths. An optional background
wavelength may also be specified and its absorbance
will be subtracted from the other wavelengths before
they are reported.
Read Blank SmartSpec Plus will zero the detector in preparation for
reading samples. It is important to zero the instrument
at each new assay. Insert a cuvette that contains all
assay components except the analyte and press Read
Blank.
Read Sample SmartSpec Plus will collect absorbance data from the
cuvette in the instrument.
Select Used to toggle between or among choices. Once the
choice is made, its selection is confirmed by pressing
Enter.
Enter Indicates to SmartSpec Plus that a choice has been
made.
14
Cancel Cancels the last entry or causes an exit from Setup.
When editing sequence information, press Cancel to
delete a base. When Options are displayed, press
Cancel to return to the assay.
and Used to access other menus or screens. Also used to
move the cursor.
Paper Feed Feeds paper through the printer. Used when removing
or installing a paper roll.
Alpha The Alpha key allows the user to toggle between
alpha and numeric input for naming stored standard
curves or naming operators. When in alpha mode, an
LED is illuminated on the key. To type an alphabetic
character when in the alpha mode, press one of the
number keys in the left or center columns. To type an
A, press the 1 key once; to type a B, press the 1 key
twice, and to type a C, press the 1 key three times.
These keys also enable the user to enter sequence data.
Sequence data are input using the last column of keys
on the number pad: to type an A, press the 3 key; to
type a C, press the 6 key; to type a G, press the 9 key
and to type a T or U, press the exp key.
Number pad Used to input numeric values for SmartSpec Plus
calculations. The exp key is used for making entries in
exponential notation, and when it is pressed, an e is
displayed by SmartSpec Plus. It refers to the base 10
and not to the base of the natural logarithm.
Section 5
Quick Guide
1. Power up the instrument. It will go through system status check and display an error
message if there are any problems.
2. Press one of the Assay buttons.
A. DNA/RNA.
i. Choose type of nucleic acid
a. dsDNA, ssDNA or RNA: accept or modify the conversion factor
b. DNA oligo or RNA oligo: input molar extinction coefficient and molecular
weight; otherwise choose method for SmartSpec Plus to estimate these values.
ii. Choose whether to subtract background and, if so, specify background
wavelength.
B. Protein
i. Choose type of assay.
a. Bradford. Measure absorbance at 595 nm.
b. Lowry. Measure absorbance at 750 nm.
c. BCA. Measure absorbance at 562 nm.
d. UV. Measure absorbance at 260, 280, and 320 nm.
e. Other. User specifies the wavelength to read.
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