BIO RAD SmartSpec 3000 Instruction Manual Download Page 1

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SmartSpec

™

3000

Spectrophotometer

Instruction Manual

Catalog Number

170-2501

For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)

Summary of Contents for SmartSpec 3000

Page 1: ...SmartSpec 3000 Spectrophotometer Instruction Manual Catalog Number 170 2501 For Technical Service Call Your Local Bio Rad Office or in the U S Call 1 800 4BIORAD 1 800 424 6723...

Page 2: ...Section 8 Description of Assays 23 Section 9 Troubleshooting 46 9 1 Troubleshooting Table 46 Section 10 Maintenance 47 10 1 Cleaning Up a Fluid Spill 47 10 2 Cleaning Quartz Cuvettes 47 10 3 Changing...

Page 3: ...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 1 safety standards Certified products are...

Page 4: ...Do not use near flammable materials 2 Always inspect the SmartSpec 3000 for damaged components before use 3 Always connect the system to the correct AC power source 4 Always connect the correct instru...

Page 5: ...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 t...

Page 6: ...h 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 Vi...

Page 7: ...e 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 Initi...

Page 8: ...nce of a sample Fig 5 A symbolic representation of the SmartSpec 3000 optics module The path of light in the SmartSpec 3000 is as follows Light from the xenon flash lamp Lens Sample Slit Concave mirro...

Page 9: ...T log 10 I0 It log10 10 cl cl Where I0 Intensity of incident light It Intensity of transmitted light Molar extinction coefficient liter mole cm c Sample concentration mol liter l Optical pathlength Fi...

Page 10: ...od to determine DNA and RNA oligonucleotide molar extinction coefficient and molecular weight 5 Default number of sample replicates 6 New operator name 7 Current operator 8 Serial port export 9 Curren...

Page 11: ...and molar pmol l concentrations You can input the molecular weight and molar extinction coefficient directly if you know them otherwise SmartSpec 3000 will use one of three methods to estimate them f...

Page 12: ...t export When this feature is activated absorbance data and concentration data are sent out through the serial port in all assays Otherwise only the Scan and Kinetic assay data can be sent out the ser...

Page 13: ...onnecting the SmartSpec 3000 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...

Page 14: ...f 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...

Page 15: ...me 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 exporte...

Page 16: ...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 3000 calculations The exp key is used for making entries in...

Page 17: ...ckground and if so specify background wavelength 3 If your dilution factor is not 1 0 set the dilution factor 4 Zero the instrument Place a cuvette containing the blank solution into SmartSpec 3000 an...

Page 18: ...ersion and dilution factors Concentration A260 Conversion Factor Dilution Factor or Concentration A260 A320 Conversion Factor Dilution Factor SmartSpec 3000 is programmed with default conversion facto...

Page 19: ...ve The instrument will report the slope and intercept of the standard curve and the square of the correlation coefficient for the curve r2 The correlation coefficient r provides an indication of how w...

Page 20: ...pecify more than one repetition for slow scanning Scan data may be downloaded to an external computer via the serial port or may be printed by SmartSpec 3000 D Kinetic SmartSpec 3000 can collect absor...

Page 21: ...ber greater than or equal to 1 if one volume of sample has been diluted with nine volumes of buffer the dilution factor is entered as 10 Use the number keys to input the dilution factor and press Ente...

Page 22: ...st going to the Ready screen if absorbance data are displayed you can return to the Ready screen by pressing Enter Press the right arrow for Options Press the right arrow key to change the number of s...

Page 23: ...rint to begin printing the scan If you do not want to print the scan press Cancel From this screen you can press the left arrow key to select nm scale and direct SmartSpec 3000 to print only a subset...

Page 24: ...he data are printed the local minimum and maxi mum in each region are printed Smoothing will take a running average of five consecutive data points two on either side of the current wavelength and plo...

Page 25: ...sample by placing the cuvette in the chamber and pressing Read Sample You can also return to the Ready screen by pressing Enter Available options are Change next sample number and Change number of sam...

Page 26: ...o signify your selection 2 Determining the conversion factor A dsDNA ssDNA or RNA If you choose dsDNA ssDNA or RNA the next screens display the conversion factor and allow you to modify it if you want...

Page 27: ...Note You must enter the molar extinction coefficient in units of liter mole 1 cm 1 SmartSpec 3000 will next prompt for the oligo molecular weight If you know the molecular weight press Select to togg...

Page 28: ...s absorbance conversion factor SmartSpec 3000 will estimate them for you If you know the conversion factor then press Select to toggle from No to Yes and press Enter If you do not know the conversion...

Page 29: ...ethods it will use length composition or sequence The calculated molar extinction coefficient and molecular weight will be displayed at the end A fourth option of Do not need may be selected if you do...

Page 30: ...and to convert those absorbance data into concentrations of g ml and pmol l c Sequence SmartSpec 3000 will automatically shift to the Alpha mode on the keypad so that you enter an A by pressing the 3...

Page 31: ...be read Use the number keys to enter the desired reading wavelength and press Enter 2 Background subtraction You will next be offered a chance to choose a wavelength for a background reading If you do...

Page 32: ...you that it will not be able to convert absorbance readings into concentrations If you want SmartSpec 3000 to convert absorbance to concentration press Select to toggle from Yes to No and press Enter...

Page 33: ...previous screen by pressing the left arrow key Press 0 to see the name of the standard curve and the date that it was created Return to the assay by pressing the right arrow key or go back to the prev...

Page 34: ...and press Enter Next SmartSpec 3000 will prompt for the units of concentration Your standards must all have the same units Press Select until you see the desired units of concentration and then press...

Page 35: ...concentration for all standards must be the same as those specified before the first standard was read When collecting data for construction of the standard curve SmartSpec 3000 does not display the...

Page 36: ...ficant digits because SmartSpec 3000 absorbance data are measured to four significant digits Once you have selected the units of concentration SmartSpec 3000 will begin to collect absorbance measureme...

Page 37: ...lso be printed at the end of the assay Press the left arrow key to view the absorbance of the last replicate or Press the right arrow key to view the mean and standard deviation of the absorbance of c...

Page 38: ...d and then for units of concentration Use the number keys to input the concentration of standard 1 and press Enter You may enter concentrations with as many as eight significant digits but all results...

Page 39: ...ion of the current replicate group All standard absorbance data can also be printed at the end of the assay Press the left arrow key to view the absorbance of the last replicate or Press the right arr...

Page 40: ...until the standard you want to edit is displayed and then press Enter Use the number keys to input the correct concentration and press Enter Press the left arrow key to return to the assay and comple...

Page 41: ...ulations may not be very accurate At the end of the assay you will be offered a chance to save the standard curve into memory for future use either as a standard curve or to compare to another standar...

Page 42: ...t a standard curve when it is first created or when it is recalled from memory When a standard curve is newly created the following prompt is displayed Press Select to toggle from No to Yes and press...

Page 43: ...maximum Y value is automatically calculated and the graph is plotted When you rescale the Y axis you must input the new maximum Y value and press Enter A new maximum X value is automatically calculate...

Page 44: ...sorbance readings at 255 256 257 258 and 259 nm and plot this average at 257 Then it will average the absorbance readings at 256 257 258 259 and 260 and plot this average at 258 nm This is available t...

Page 45: ...ose 0 if you don t want any subregions If you specify 1 or more subregions then you are next prompted to define them Use the number keys to input the beginning wavelength and press Enter Then use the...

Page 46: ...No to Yes and press Enter Use the number keys to input the background wavelength and press Enter SmartSpec 3000 is now ready to collect kinetic absorbance data at the specified wavelength For best res...

Page 47: ...erwise press Select to toggle from Yes to No and press Enter Use the number keys and the exp key to input the new conversion factor and press Enter SmartSpec 3000 is now ready to collect OD600 reading...

Page 48: ...measurement is subtracted from all readings If you do not want to subtract a background reading press Enter and SmartSpec 3000 will be ready to collect absorbance data If you want to subtract a backg...

Page 49: ...sors over Paper feed button keyboard 2 Call Bio Rad service is faulty Print out text too dark Printer paper damaged 1 Replace with new paper roll caused from excessive heat moisture Paper loaded incor...

Page 50: ...ple is in the recommended working range i e 0 1 1 5 AU Section 10 Maintenance 10 1 Cleaning Up a Fluid Spill In case of a spill in the cuvette chamber clean up the spill using the following procedure...

Page 51: ...oll as shown Do not fold or crease leading edge of paper Feed paper into slot opening until it stops Press the Paper Feed key located on the keypad and guide the paper into the printer until it grabs...

Page 52: ...nds max Photometric repeatability 0 02 AU in a 0 1 2 0 AU range 9 readings and over a 200 800 nm range Dimensions 33 cm W x 26 cm D x 14 cm H Weight 3 6 k g Power requirement 90 260 VAC 47 63 Hz Regul...

Page 53: ...Factor Dilution Factor g ml concentration OR A260 A320 Conversion Factor Dilution Factor g ml concentration Default conversion factors are 1 0 O D unit 50 g ml dsDNA concentration 1 0 O D unit 40 g m...

Page 54: ...170 2507 Spring cuvette holder 12 2 Bio Rad Protein Assays Catalog Number Product Description 500 0001 Bio Rad Protein Assay Kit I 450 standard assays bovine gamma globulin standard based on the meth...

Page 55: ...rects for pathlengths shorter than 10 mm when converting absorbances into concentration The following table gives you the absorbance at A260 and corresponding concentration for the different pathlengt...

Page 56: ...y 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 HyperTe...

Page 57: ...ile 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...

Page 58: ...47 95 69 65 Fx 01 47 41 9133 Israel Italy Japan Korea The Netherlands Ph 47 23 38 41 30 Fx 47 23 38 41 39 Sweden Ph 46 0 8 55 51 27 00 Fx 46 0 8 55 51 27 80 Switzerland Ph 061 717 9555 Fx 061 717 955...

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