Bio-rad SmartSpec Plus Spectrophotometer User Manual Page 1

SmartSpec™PlusSpectrophotometerInstruction ManualCatalog Number170-2525For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-

4.2 SetupWhen the SmartSpec Plus is not carrying out an assay, you can enter the Setup routineby pressing Setup. To proceed through Setup without cha

3. Background compensation. When using the DNA/RNA assay, you can choose tohave SmartSpec Plus automatically subtract background absorbance readings

7. New Operator. When SmartSpec Plus prints a report, it includes the name of theoperator (if specified). You can add a new operators name to the li

12. Time convention. You may choose to print time in 12-hour or 24-hour mode. Togglebetween the choices by pressing Select. and then press Enter. Pre

The serial port parameters are: 9600 baud rate, 8 data bits, no parity, 1 stop, andXON/XOFF flow control. These parameters are pre-set in the unit. Wh

4. When the SmartSpec Plus sends data, either when requested from Scan, or Kinetics, orwhen the user activates the data echo toggle in Setup, the data

4.5 Keypad DescriptionsPrint SmartSpec Plus will print data on the current sample,a user-selectable range of samples or all samples in thecurrent ass

Protein SmartSpec Plus will automatically read one or morewavelengths depending on the type of assay specified(Bradford, Lowry, BCA or UV Protein) or

Cancel Cancels the last entry or causes an exit from Setup.When editing sequence information, press Cancel todelete a base. When Options are displayed

ii. Choose standard curve option.a. Create a new standard curve.b. Recall a standard curve from memory.c. No standard curve. SmartSpec Plus will not b

Table of ContentsPageSection 1 General Safety Information ...11.1 Caution Symbol...

Section 6Operating InstructionsNote: It is Good Laboratory Practice (GLP) to use the SmartSpec Plus spectrophotometerto measure absorbances in the ra

The SmartSpec Plus Bradford, Lowry, BCA and Other Protein assays use standardcurves created by measuring the absorbances of solutions of known concent

Limitations of the methods. The linear regression, when fit properly, generallyproduces reliable results at the extremes of the range of the standard

2. Choose sample replicates. Samples may be read in replicate and you do not need tohave the same number of replicates for each different sample. For

• Press the right arrow key to change the number of sample replicates.B. Sample replicates.• Place the first replicate of your first sample in the cuv

• Press A260:A280 to see the ratio displayed• For DNA oligos and RNA oligos, press Conc to see the concentration displayed inunits of pmoles/µl.• To r

• Now press Select and the negative sign will be displayed in front of 0.500.• Press Enter again and now define the upper limit. The upper limit must

• If you want to download the data, make sure the connections are made to thecomputer and then press Enter. If you do not want to download the data, p

8. Exit the Assay. You can exit the assay by pressing the left arrow key when the Readyscreen is displayed.SmartSpec Plus will prompt you to save a ne

• After the default conversion factor is accepted or changed, SmartSpec Plus is readyto measure nucleic acid absorbance and convert that absorbance to

WarrantyThe SmartSpec Plus Spectrophotometer and accessories are warranted against defectsin materials and workmanship for one year. If any defects oc

b. If you do not know the molecular weight.• Use the number keys to input the number of A’s, then press Enter to move thecursor to the next field and

• Press > to continue the assay, and the screen will display• SmartSpec Plus is now ready to collect absorbance data.b. If you do not know the mole

• Use the number keys to input the length of the oligo and press Enter. You canenter a length of up to 99 nucleotides. After you press Enter, SmartSpe

• Press the right arrow to continue the assay. SmartSpec Plus will now be readyto measure the absorbance of your oligonucleotide and to convert thosea

7.2 Protein Assay1. Choosing the type of assay. Press Protein and the display becomesAs you press Select, other options are displayed:Press Enter whe

3. Standard Curves. Once you have selected the type of assay, SmartSpec Plus will askif you want to make a new standard curve. You can decline to mak

• In order to see the information about the standard curve, press Select to togglefrom No to Yes and press Enter.r2 refers to the square of the correl

• Each time you place a new blank into SmartSpec Plus, press Read Blank.After the absorbance reading is collected, the instrument is zeroed and the di

i. If there are no standard replicates. SmartSpec Plus will direct you to enterthe concentration of each standard and then to measure the absorbance o

• If you have varying numbers of replicates for the different standards, pressSelect to toggle from Yes to No and press Enter. Skip ahead to b. If you

Section 1General Safety InformationThis instrument is intended for laboratory use only.This product conforms to the "Class A" standards for

After all replicates of standard #1 are measured, SmartSpec Plus asks for the concentrationof standard #2 and directs you to put the replicates into S

b. If there are varying numbers of standard replicates. Each time SmartSpec Plusprepares to read a standard, you will be prompted for the number of r

After all replicates of standard #1 are measured, SmartSpec Plus asks for the number ofreplicates of standard #2 and then the concentration of standar

The display will show eitheror• Press the right arrow key to return to the assay.After the last replicate of the last standard is measured, SmartSpec

• You can choose to blank the instrument again before you re-read your sample bypressing Enter at this screen. If you do not want to re-zero SmartSpec

• Press 0 to see the intercept. Press the right arrow to return to the assay or press theleft arrow key to see the slope displayed again.If the slope

If you want to rescale the Y axis, press Enter. If you want to rescale the X axis, pressSelect to toggle the display and press Enter.When you rescale

7.3 Spectral ScanningSmartSpec Plus can collect data over a range of wavelengths and then plot thoseabsorbances. There are both Fast and Slow scans:

• Use the number keys to input the number of successive scans and press Enter.SmartSpec Plus will now display the Ready screen. If you want to invoke

7.4 KineticsWhen you press Kinetic, you begin by specifying the reading wavelength.• Use the number keys to input the wavelength to be read and press

Section 3Installation3.1 Environmental RequirementsTo ensure correct operation and stable performance over an extended period of time,install the Sma

7.5 OD600This assay is useful for monitoring growth of bacterial cultures. When you press OD600,SmartSpec Plus will display the current conversion fa

7.6 Lambda λλThis assay allows you to collect absorbance data from up to three different wavelengthssimultaneously. No concentrations are calculated.

Section 8TroubleshootingTroubleshooting TableProblem Cause SolutionUnit does not power up Power switch in off position. 1. Check power switch on back

Problem Cause SolutionRS232 serial port/no Incomplete system 1. Power down unit/Restart unit.response initialization during power up.PC setup incorrec

4. Dampen cloth with water. Use it to clean the instrument from the outside, including thedisplay and the keyboard.9.2 Cleaning Quartz CuvettesIt is

• Trim leading edge of paper roll with scissors if uneven or torn.• Feed paper into slot opening until it stops. Press the "Paper Feed" key

Section 11System, Accessories, and Reagents for theSmartSpec Plus11.1 Product InformationCatalogNumber Product DescriptionSmartSpec Plus Spectrophoto

11.3 Cuvette Selection GuideCatalog Minimum Maximum CuvetteNumber Volume Volume Type Pathlength170-2502 1,000 µl 3,500 µl Standard cuvette, quartz 10

Life ScienceGroup Sig 0603Bio-RadLaboratories, Inc.Web siteUSA(800) 4BIORAD Australia02 9914 2800 Austria(01)-877 89 01 Belgium09-385 55 11 Braz

Section 4Introduction4.1 Identification of System ComponentsFig. 1. View of the SmartSpec Plus Spectrophotometer.Instrument1. Keypad interface Chang

Fig. 3. View of the SmartSpec Plus sample compartment.9. Cuvette chamber Insert cuvette into the cuvette chamber. Ensure thatthe cuvette is placed in

5UV/Visible Spectroscopy is based on the absorption of light as a function of wavelength.All spectrophotometers have a light source that generates lig

The Lambert-Beer’s law identifies the relation between sample concentration and theintensity of transmitted light. Absorbances and % transmittances ar