biochrom Ultrospec 3300 pro, Manual

Page 1: ...patibility Generic emission standard part 1 Electrical equipment for measurement control and laboratory use EN 61000 4 6 1992 Electromagnetic compatibility Generic immunity standard part 1 Residential...

Page 2: ...tilities 33 Output to Printer 35 Download to Spreadsheet 36 Messages 36 ACCESSORIES 38 Multiple Cell Holder Accessories 38 Single Cell Holder Accessories 39 Other Accessories consumables etc 40 SWIFT...

Page 3: ...works see Operation It can be configured to have the display and print outs in English 0 German 1 French 2 Spanish 3 Italian 4 or Russian 5 by pressing the number in brackets as the instrument powers...

Page 4: ...symbols and their meaning Caution refer to accompanying documents Background colour is yellow symbol and outline are black UV RADIATION UV RADIATION IS HARMFUL TO YOUR EYES HOT If power is restored wi...

Page 5: ...ses through your cell containing the sample of interest and then a defocusing lens to a solid state detector unit The resulting signal is then filtered and displayed Your spectrophotometer Measures st...

Page 6: ...l wavelengths in the mode selected this is subtracted from all subsequent samples in the experiment press print to output graphics and results to a parallel printer or PC this is automatic if Auto pri...

Page 7: ...rmation presented in a box on the LCD b Other measurement modes have a graphical layout with information about the instrument also being provided along a status bar which shows messages whether a prin...

Page 8: ...is used for subsequent samples until changed Insert samples as required and press run repeat as necessary To go back and change the wavelength press mode Transmission Transmission mode measures the a...

Page 9: ...alent to a one point calibration and assumes that sample of zero concentration has zero absorbance The procedure is as follows Enter appropriate wavelength and press enter Enter appropriate sample num...

Page 10: ...mory 1 Press C 2 Select whether it is between 1 10 11 20 etc by using 4 3 Select the method number 4 Confirm yes enter or no 4 enter 4 Save Stored methods up to a maximum of 50 are saved directly from...

Page 11: ...Extracting nucleic acids from cells is accompanied by protein and extensive purification is required to separate the protein impurity The 260 280 ratio gives an indication of purity it is only this h...

Page 12: ...o modes Use of 7 l volume ultramicrovolume cell If using this cell please note that it has a 5 mm pathlength and that these modes assume a 10 mm pathlength cell is used Ensure the cell is filled corre...

Page 13: ...account for potential differences between cells UViMicro cells are ideal for this application since the minimum volume that can be used is 20 l If selected enter wavelength required Default values ar...

Page 14: ...ation is still possible see Results below The results for cDNA concentration ng l dye probe concentration pmo l and nucleotides dye are presented Results Use this facility to view the underlying absor...

Page 15: ...using 4 This is very useful for optimisation note that concentration and ratio results are updated automatically Tm Calculation Tm the theoretical annealing temperature can be calculated for a primer...

Page 16: ...e colour obtained from the reaction of Folin Ciocalteu phenol reagent with the tyrosyl residues of an unknown protein and comparing this colour value to those derived from a standard curve of a standa...

Page 17: ...________________________________________________________________ ___ Issue 02 07 2001 English 17 Information Information relating to proteins and amino acids...

Page 18: ...line fit are calculated see Appendix Linear interpolation joins up consecutive data points by a series of straight lines Spline calculates and fits the best curved line through the data points using...

Page 19: ...andards insert reference and standards and select yes If loading samples insert reference and samples and select no Samples have to be run separately and individually If a sample absorbance is within...

Page 20: ...he two coefficients In cases where Factor 2 is found to be negative it should be set to zero since it means there is no contribution to the protein concentration due to absorbance at 260 nm The use of...

Page 21: ...hod if required using 4 Insert reference and samples and press run 215 225 nm method Protein concentration g ml can be determined using the empirical formula below Waddell W J 1956 A simple UV spectro...

Page 22: ...both multiple peaks and inflections at the same time The procedure is as follows Set up Enter start wavelength range 190 1090nm Enter end wavelength range 200 1100nm Select scan speed as appropriate...

Page 23: ...and printing purposes by using scale and offset to affect the selected data type Note that different individual scans cannot be overlaid Select data type as appropriate off 1st or 2nd or 4th derivativ...

Page 24: ...elect units using 4 Save method if required using 4 Insert reference and samples and press run Abs Diff This facility enables the determination of the following equations 1 Abs 1 Abs 2 Factor 1 for bi...

Page 25: ...e first wavelength this is wavelength on the UV side of the peak Enter the second wavelength this is the wavelength of the peak and will have a value greater than the first wavelength Enter the third...

Page 26: ...integration times for very low and very high absorbance readings Factors Enter the overall dilution factor C Enter the first factor Maximum of 2 decimal places Press C on the keypad to enter a negativ...

Page 27: ...s selected Select if active reference is required using 4 Use if reference changes with time and needs to be subtracted Select time units seconds or minutes using 4 Enter the delay time over which no...

Page 28: ...is required post run Select if data points should be indicated on each assay Select if the graph should be printed with the results Post Run This facility enables an assay to be selected in serial or...

Page 29: ...a points by a series of straight lines Spline calculates and fits the best curved line through the data points using a natural cubic spline fitting method requires a minimum of 4 data points Set up En...

Page 30: ...andards insert reference and standards and select yes If loading samples insert reference and samples and select no Samples have to be run separately and individually If a sample absorbance is within...

Page 31: ...s to be applied to a change in absorbance with time use the kinetics application Set up Enter the wavelength Select the curve type from linear regression linear interpolation spline using 4 Enter the...

Page 32: ...andards insert reference and standards and select yes If loading samples insert reference and samples and select no Samples have to be run separately and individually If a sample absorbance is within...

Page 33: ...ntrast option to make the background white GLP This facility is only available if the GLP option in Set up has been enabled for further information on GLP refer to the Appendix The GLP Calibration Res...

Page 34: ...re printed out every time run is pressed Select between output to printer only output to computer only output to printer and computer using 4 Select if automatic print out of parameters and graph if i...

Page 35: ...ter set Epson FX 80 Epson 9 pin Includes Epson FX 850 and similar Epson 24 pin ESC P For use with Epson 24 pin dot matrix printers and older inkjet printers such as the Stylus 400 Epson InkJet ESC P2...

Page 36: ...o hard disk Results from all modes of use on the instrument can be output in this way Output is automatic when print key is pressed Messages Most messages are self explanatory and relate to use of the...

Page 37: ..._____________________ ___ Issue 02 07 2001 English 37 Beam blocked Not enough light getting to detector check light beam is not blocked by a cell 3 0 Sample too concentrated or something blocking ligh...

Page 38: ...have the option of being used as a single cell holder This means that there will be no rotation after pressing run Description Part number Comments 4 position cell changer 80 2106 01 Accommodates cel...

Page 39: ...holder 80 2106 09 Use with 50 l cell 80 2076 38 Cylindrical cell holder 80 2106 10 Up to 100 mm pathlength cylindrical cells Water heated cell holder 80 2106 08 10 40 mm pathlength Requires a water ci...

Page 40: ...cell holder 80 2106 14 Printer stand 80 2112 13 For Seiko DPU 414 thermal printer Dust cover 80 2106 19 Spare Consumables and other items Pump head tubes 6 for Sipper 80 2080 74 PTFE flowcell tubing w...

Page 41: ...eral analytical purposes Wavelength Scanning Reaction Kinetics Quantification Time Drive 80 2108 31 SWIFT II METHOD for method development Wavelength Scanning Reaction Kinetics Quantification Time Dri...

Page 42: ...ers and calibrated test equipment Approved to ISO 9001 standard Choice of agreement apart from break down coverage can include Preventative maintenance Certification When using calibration standard fi...

Page 43: ...are covered by their own warranty an engineer s call out fee is not and users are advised to change their own lamps Lamp replacement is very easy and the process has been designed so that the user ca...

Page 44: ...e lamp assembly housing and tilt it upwards 6 Slide the lamp plate out and unplug the cable connector If the tungsten lamp has failed the replacement should be inserted onto the plate pushing it all t...

Page 45: ...nnect the power supply cord The fuse holder can only be opened if the power supply plug has been removed and is located in the power input socket on the back panel of the instrument 2 Slide the fuse h...

Page 46: ...ed in a chemical resistant finish Strong concentration of sample however may affect the surface and spillages should be dealt with immediately Observe all necessary precautions if dealing with samples...

Page 47: ...ment the spectrum of a 0 02 v v solution of toluene in hexane should b recorded the ratio of the absorbance at the maximum 269nm and minimum 266nm should be at least 1 5 It can be shown that this requ...

Page 48: ...lues when new the wavelength accuracy by comparing to the 656nm deuterium line the values of a built in absorbance filter compared to when the instrument was manufactured or last serviced by an accred...

Page 49: ...2 56 Calibration Full UV Visible Bandwidth 1 3 1 8nm 1 7nm PASS Wavelength 656 1nm 656 0nm PASS Absorbance at 220nm 1 763 1 781A 1 772A PASS 340nm 1 633 1 665A 1 649A PASS 500nm 1 477 1 491A 1 484A PA...

Page 50: ..._______________ ___ 50 English Issue 02 07 2001 baseline stored 14 08 00 13 38 GLP enabled causes printed results to have calibration status included GLP results can also be sent to PC for electronic...

Page 51: ...ch entry Setup No of s 2 1 511 2 720 Integration time Default Factors c 100 K1 12 26 K2 0 30 K3 0 40 press C first to get the negative sign K4 27 41 press enter repeatedly to get to end of page Equati...

Page 52: ...________________________________________________________________ ___ 52 English Issue 02 07 2001 title l litre unit will appear in print out not in title Enable equation Check the equation...

Page 53: ...ter entry Description Nickel to get alphanumeric keypad press stop after entry of name Equation press K3 already defined as 0 40 A1 already defined as absorbance at 511 nm K4 already defined as 27 41...

Page 54: ...ate at which NADH is used up by measuring the absorbance of the reaction mixture at 340 nm Since LDH is in excess this rate is directly proportional to the rate of pyruvate produced in the first react...

Page 55: ..._______________ ___ Issue 02 07 2001 English 55 where dC dt rate of change in concentration mol litre L cell path length normally 1 cm and E molar absorptivity molar extinction coefficient of the comp...

Page 56: ...s activity per unit volume IU litre or kat litre In order to simplify the calculation of enzyme activity variables such as molar absorptivity and sample volume can be combined to produce a conversion...

Page 57: ...________________________________________________________________ ___ Issue 02 07 2001 English 57 working in the IU unit but to work in microkatals the conversion factor must be divided by 60...

Page 58: ...a kinetics assay or standard curve determination is calculated from a least squares linear regression of the data The following equations are used where n is the number of data points Slope x y n xy...

Page 59: ...0 5 of absorbance value to 3 000A at 546nm Stability 0 001A per hour at 340nm at 0A after warm up tungsten lamp Stray light 0 025 T at 220nm using NaI and 0 025 T at 340nm using NaNO2 Digital output...

Page 60: ...published specification The warranty included in the conditions of supply is valid for 12 months only if the product has been used according to the instructions supplied They can accept no liability f...