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23

5061-049 REV 1.4

25-9574-75UM AE 05/2010

23

Results Screen
Step 12 
Pipette on the reference sample and lower the sampling head. If Auto-Read is 
off, press the 0A/100%T key. This will be used for all subsequent samples until 
changed. If QA is switched on the sample will need to be replaced and the 
0A/100% T key pressed again.

Step 13 
Clean the top and bottom plates, pipette on the sample and lower the sampling 
head. If Auto-Read is off, press 

. The unit measures the Absorbance and uses 

this information to calculate the Measured Tm.
Repeat step 13 for all samples.

Press 

Esc

 to return to the Life Science Screen.

Press 



 to display available Options which are described below.

Options (select using keypad numbers)
1. Return to Parameters Screen (step 1 above).
2. Print result via selected method.
7.  Sample Number – add a prefix to the sample number and reset the 

incrementing number to the desired value.

8. Save Method – use the alpha-numeric keys to enter a name for the method and 

press Save 

.

9. Auto-Print – toggles Auto-Print on/off.

Exit options by pressing 

Esc

, or wait.

4.2.6. CyDye measurement

Measurement of the labeling efficiency of fluorescently labeled DNA probes 
before 2-color microarray hybridization ensures that there is sufficient amount of 
each probe to give satisfactory hybridization signals. These data also provide an 
opportunity to balance the relative intensities of each fluorescent dye by adjusting 
the concentration of each probe before hybridization. The DNA yield is measured 
at 260nm while the incorporation of fluorescein, Cy3, Cy5 and other dyes are 
measured at their absorption peaks. This method may also be useful for measuring 
the yields and brightness of fluorescently labeled in-situ hybridization probes.

Equations used in the software are detailed below:

DNA Concentration  
= (A

260

 – A

Background

) * NucleicAcid Factor 

DNA Quantity  
= DNAConcentration * Volume

Dye Concentration  
= ((A

Dye

- A

Background

) / ExtinctionCoefficient

Dye

) * PathlengthFactor * Dilution_Factor 

* 10

6

Dye Quantity   
= DyeConcentration * Volume

Содержание NanoVue Plus

Страница 1: ...oVue Plus Product User Manual Biochrom US Telephone 1 508 893 8999 84 October Hill Rd Toll Free 1 800 272 2775 Holliston MA Fax 1 508 429 5732 01746 1388 support hbiosci com USA www biochromspectros c...

Страница 2: ...19 4 2 5 Tm Calculation 20 4 2 6 CyDye measurement 23 4 3 Proteins 26 4 3 1 Theory 26 4 3 2 Protein UV 28 4 3 3 Protein A280 30 4 3 4 BCA 31 4 3 5 Bradford 34 4 3 6 Lowry 36 4 3 7 Biuret 39 5 APPLICA...

Страница 3: ...e user with basic use troubleshooting and how to use the instrument in a safe manner CAUTION This instrument contains a UV source which generates a light beam that is present at the sample platform Do...

Страница 4: ...t is used in a manner not specified or in environmental conditions not appropriate for safe operation the protection provided may be impaired and instrument warranty withdrawn There are no user servic...

Страница 5: ...he general software features of the instrument including wavelength scanning single or multi wavelength Absorbance and concentration measurement kinetics standard curves and Absorbance ratio can be ut...

Страница 6: ...cience applications such as nucleic acid and protein assays 2 Applications General spectroscopic applications 3 A folder to store your more frequently used methods Inactive when empty 4 Methods Contai...

Страница 7: ...heir normalized 10 mm pathlength values to allow the use of literature based factors for concentration measurements Within the DNA RNA Oligonucleotide Tm Calculation Cy Dye Protein UV and Protein A280...

Страница 8: ...y out a new reference scan at any time by pressing the 0A 100 T key prior to lowering the sampling head or to perform a series of readings or reference scans without raising the sampling head 2 7 Qual...

Страница 9: ...00 A or 100 T on a reference solution at the current wavelength in the mode selected When in scan mode make a reference scan Confirm selection Confirms a selection Take measurement Makes a measurement...

Страница 10: ...to enter the Options menu 2 9 Software style The user interface is built around folders of files which are displayed on the first page when the instrument is switched on Different folders are numbered...

Страница 11: ...Filtration may also be advantageous Step 2 Carefully apply the sample so that it sits over the black spot between the four alignment spots see Figure 4 Take care not to introduce bubbles into the samp...

Страница 12: ...lace them as a pair The glass plates are permanently fixed within plastic housings which clip to the sample handling module Replacements are available from your supplier Exchanging the sample plates i...

Страница 13: ...ckwise and turn the green knob manually Step 4 Replace the cover Figure 8a Figure 8b 25 9574 75UM AE 05 2010 13 3 3 Cleaning and general maintenance Before cleaning the case of the instrument switch o...

Страница 14: ...protein determination Pressing 6 enters the Protein sub folder which contains various standard methods for protein determination 4 2 Nucleic Acids 4 2 1 Theory Nucleic acids can be quantified at 260...

Страница 15: ...l result It is advisable to ensure that the Abs260 value is greater than 0 1 A for accurate measurements An elevated Absorbance at 230 nm can also indicate the presence of impurities 230 nm is near th...

Страница 16: ...lution Factor Screen see second parameter screen to the left Enter the Volume of the sample using the keypad numbers range 0 01 to 9999 Press the down arrow Enter the volume of the Diluent using the k...

Страница 17: ...n plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if Background Correction selected 320 nm 7 Sample Number add a prefix to the sample number and reset the incrementing num...

Страница 18: ...ean the top and bottom plates pipette on the sample and lower the sampling head If Auto Read is off press This measures at the selected wavelengths and displays the results The ratio of the Absorbance...

Страница 19: ...t arrows Press the down arrow Step 5 Select the Units of measurement using the left and right arrows Options g ml ng l g l and pmol l If pmol l is selected the factor changes to a selection table deno...

Страница 20: ...relation to its neighbor Breslauer et al Proc Natl Acad Sci USA Vol 83 p3746 1986 The data obtained are useful both in the characterization of oligonucleotides and in calculating Tm for primers used...

Страница 21: ...eight ABCDE EABCDE where EABCDE 2 EAB EBC ECD EDE EB EC ED The molecular weight MW of a DNA oligonucleotide is calculated from MW g mole dA 312 2 dC 288 2 dG 328 2 dT 303 2 MW counter ion length of ol...

Страница 22: ...in moles l using the keypad numbers range 0 000 to 10 Press the down arrow Step 6 Select the Counter Ion Na K TEA or Other Step 7 if Other selected see the previous section Enter the molecular weight...

Страница 23: ...Method use the alpha numeric keys to enter a name for the method and press Save 9 Auto Print toggles Auto Print on off Exit options by pressing Esc or wait 4 2 6 CyDye measurement Measurement of the l...

Страница 24: ...y Enter a name for the dye to identify it correctly To complete the entry press Next to enter the second dye detail entry screen OR Press Cancel Esc to return to the Life Science Screen Step 7 If ther...

Страница 25: ...appropriate factor If a different factor is desired select Custom and the Factor can be user defined Press OK to enter the Results screen OR Press Esc to cancel the selections and return to the Param...

Страница 26: ...nt effect due to strong nucleotide Absorbance at 280 nm This can be compensated by measuring Abs260 and applying the equation of Christian and Warburg for the protein crystalline yeast enolase Biochem...

Страница 27: ...nm this is usually BSA bovine serum albumin The Biuret method depends on reaction between cupric ions and amino acid residues in an alkali solution resulting in the formation of a complex absorbing a...

Страница 28: ...g the left and right arrows Options are 0 5 mm 0 2 mm or Automatic Press the down arrow Step 3 Dilution Factor known Enter the Dilution Factor using the keypad numbers range 1 00 to 9999 Use the C but...

Страница 29: ...d 280 nm wavelengths and displays the Protein concentration as the Result Repeat step 10 for all samples Press Esc to return to the Protein Screen Press to display available Options which are describe...

Страница 30: ...n is to be used or not with the left and right arrows Press the down arrow Step 6 Mode Christian Warburg BSA IgG Lysozyme Select the Units of measurement using the left and right arrows Options mg ml...

Страница 31: ...method is fixed at 562 nm Step 3 Enter the number of Standards 1 9 to be used in the curve using the keypad numbers or left and right arrows Press the down arrow Step 4 see second image Units The user...

Страница 32: ...measured or entered using the keypad numbers if calibration mode is Manual Step 11 Standards selected Pipette on the reference sample and lower the sampling head If Auto Read is off press the 0A 100...

Страница 33: ...o 9999 Use C to backspace and clear the last digit entered and the up and down arrows to move between boxes Pressing the down arrow from the last standard will bring up the OK box Press OK to accept t...

Страница 34: ...s Next to move on to the next Parameters Screen or Esc Cancel to return to the Protein Screen Step 6 Enter the type of curve fit Options are Regression straight line Zero Regression forces the straigh...

Страница 35: ...Manual selected When all standards are measured the OK box appears Press OK to accept the calibration and go to the Results Screen see below OR Press Back Esc to cancel selections and return to the S...

Страница 36: ...the Protein Screen Press to display available Options which are described below Options select using keypad numbers 1 Return to Parameters Screen step 1 above 2 Print result via selected method 3 Togg...

Страница 37: ...rotein Screen Standards Screen Step 9 Standards Manual selected Enter the concentration values by using the keypad numbers and the up and down arrows to move between the different standard boxes range...

Страница 38: ...ows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 13 Standards Manual selected Press Next to accept the calibration and go to the Res...

Страница 39: ...g L mg l g l U l ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows the number of displayed decimal points DP to be selected from 0 to 2 Note that the resu...

Страница 40: ...ll display the results and the fitted curve as the measurements are made Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous...

Страница 41: ...ng head If Auto Read is off press the 0A 100 T key This will be used for all subsequent samples until changed If QA is switched on the sample will need to be replaced and the 0A 100 T key pressed agai...

Страница 42: ...values reported are NOT nomalized to a 10 mm pathlength so if comparison is being made to other systems the appropriate factor should be used to convert them 20 for 0 5 mm pathlength 50 for 0 2 mm The...

Страница 43: ...reen The Result at the selected wavelength is displayed on the screen Use the left and right arrows to move the cursor and display the value at the cursor position 15 nm from set wavelength Press Canc...

Страница 44: ...thlength if needed Step 5 if Standard is selected Enter the Concentration using keypad numbers range 0 01 9999 Use the C button to delete the last digit entered Press the down arrow key Step 6 Units T...

Страница 45: ...p 9 Clean the top and bottom plates pipette on the sample and lower the sampling head If Auto Read is off press The concentration of the sample is displayed Results shown as indicate that the concentr...

Страница 46: ...selections and return to the Applications Screen by pressing Cancel Esc Step 6 Pipette on the reference sample and lower the sampling head If Auto Read is off press the 0A 100 T key This will be used...

Страница 47: ...to be detected Minimum Pk Width Minimum width of the peak as determined by the difference in wavelength between the higher of the two adjacent minima and the opposing intersection of that higher mini...

Страница 48: ...d can be used to provide useful information when an appropriate factor defined in the reagent kit protocol is applied Reaction rate and enzyme activity can be calculated if the factor used takes accou...

Страница 49: ...will always be fixed to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen param...

Страница 50: ...Auto Print toggles Auto Print on off Exit Options by pressing Esc or wait 5 6 Standard Curve The construction of a multi point calibration curve from standards of known concentration to quantify unkno...

Страница 51: ...rds Screen OR Press Cancel Esc to cancel selections and return to the Applications Screen Standards Screen Step 8 Enter the concentration values by using the keypad numbers and the up and down arrows...

Страница 52: ...arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous reading Step 12 Press Next to accept the calibration and go to the Results Screen see below O...

Страница 53: ...A is to be expected The procedure is as follows Step 1 Select the number of Wavelengths range 2 5 Press the down arrow Step 2 Select Pathlength from 0 5 mm or 0 2 mm Press to go to the second Paramete...

Страница 54: ...easurements on samples measuring the amount of light that has passed through a sample relative to a blank this can be air at two wavelengths Note that the individual absorbencies are not nomalized to...

Страница 55: ...ll need to be replaced and the 0A 100 T key pressed again Step 11 Clean the top and bottom plates pipette on the sample and lower the sampling head If Auto Read is off press Repeat Step 11 for all sam...

Страница 56: ...cel Esc to return to the Results Screen without storing the settings Saved Methods can be locked unlocked and deleted using the Options menu Go to the relevant folder by selecting either option 3 Favo...

Страница 57: ...arrows Press the down arrow Enter the Month Press the down arrow Enter the Year Press the down arrow Enter the Hour Press the down arrow Enter the Minute Seconds are zeroed when OK is pressed Press OK...

Страница 58: ...mat default or a list 3 History selects whether to use previously entered parameters when the instrument is switched on or whether to use defaults 4 Auto Standby selects whether to use a standby mode...

Страница 59: ...ary Press the down arrow Step 7 Press OK to enter the Results Screen and begin making measurements OR Esc to return to the Life Science Screen Step 8 Pipette on the reference sample of deionised water...

Страница 60: ...proceed to Step 4 Step 4 The Calibration process now requires 10 measurements of the calibration fluid to construct a statistically valid result The consistency of the results can be seen as the table...

Страница 61: ...oubleshooting Section 9 to adjust the pathlengths 2 The standard deviation for the 10 measurements is 1 5 If this happens repeat steps 7 7 clearing any previous results If the instrument has a Printer...

Страница 62: ...enter a number Invalid numbers cannot be entered Cells can be locked or unlocked by using the decimal point Unlocked cells can be cleared using the C key see also Option key below The user mode start...

Страница 63: ...module 8 1 Printer installation Step 1 Remove the power cable and turn the instrument over onto a soft surface taking care not to damage the sampling head Release the outermost screws using the Allen...

Страница 64: ...15 Figure 16 Figure 17 Replace the top cover plate invert the instrument and replace the cap head screws Switch the instrument on and go to utilities instrument preferences and select the Built in pri...

Страница 65: ...oth module Step 4 Note the slots in the base of the case The two lugs on the Bluetooth module plug into these Step 5 Load the rear cover into the two slots provided note the large flange is to face up...

Страница 66: ...t carefully and replace the cap head screws Step 8 Switch the instrument on and go to the preferences page under utilities instrument and select the Bluetooth option Figure 21 Figure 20 Step 5 Load th...

Страница 67: ...d Step 2 Turn the instrument back over and lift the accessory cover vertically upwards to remove Remove the tie wrap from the cable Step 3 Plug the accessory cable into the SD memory card PCB module S...

Страница 68: ...al short cut Press Save Method Use the right and left arrows to select the folder on the SD memory card to which you wish to save the method Change the filename if required Press Save NOTE A maximum o...

Страница 69: ...l way Removing the card whilst the LED is on will corrupt the collected data set All results are stored as an individual file in a directory called Serial no PVC on the SD memory card this directory s...

Страница 70: ...alibration certification Certificated engineers and calibrated test equipment Approved to ISO 9001 standard Choice of agreement apart from break down coverage can include Preventative maintenance Cert...

Страница 71: ...5 l of distilled water onto the correct position on the sample plate Close the sampling head clear all previous readings using the cursors and C key Press the 0A 100 T key as in section 7 7 This will...

Страница 72: ...technical support department of your supplier What is it What should you do Cell holder obstructed no light Check that you are using the correct power supply It should be an 18 V unit Make sure the sa...

Страница 73: ...the instrument disconnect and reconnect the power supply and then turn on the instrument If the problem persists please contact your supplier Calibration problem Check that you are using the correct p...

Страница 74: ...r calibration You may be keeping you finger on the ON OFF button too long so that the instrument receives both ON and OFF signals and switches off after the calibration Try adjusting the timing of you...

Страница 75: ...ect the accuracy of the measurement due to their volatility are therefore nor recommended Q Why does the reference blank require 2 samples A To set the reference the software will ask for 2 separate s...

Страница 76: ...e as an Excel spreadsheet Excel must be installed on the PC Extensible Meta Format an EMF graphics file a comma delimited csv data file a tab delimited txt file Rich Text Format RTF or in native PVC f...

Страница 77: ...4 5 kg Power input 18 Vdc from a 90 250 V 50 60 Hz Max 30 VA mains power pack Specifications are measured after the instrument has warmed up at a constant ambient temperature and are typical of a pro...

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