SPECTRO-NANO - MRCLAB · Operation Manual SPECTRO-NANO ... Step5: After installation, ... Language - Select operation language. Factory set: Factory default setting

Operation ManualSPECTRO-NANO

Micro-Spectrophotometer

PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION

3, Hagavish st. Israel 58817 Tel: 972 3 5595252, Fax: 972 3 5594529 [email protected]

MRC. 4.18

Foreword

Thank you for purchasing a Micro-Spectrophotometer. This manual

provided as operational and easy troubleshooting guide. Please read this

instruction carefully before operation and save for future reference.

Opening CheckPlease check the instrument and Appendix with the packing list when you

first open the package. If there is anything don’t match, please contact with

the vendor.

Safety Warnings and Guidelines1 Warning

2 Safety TipsThe operation, maintenance and repair of the instrument should comply with thebasic guidelines and the remarked warning below. If you don’t comply with them, itwill have effect on the scheduled using life of the instrument and the protectionprovided.

3 The maintenance of instrumentThe pedestal should be cleaned by the cloth stained with pure water.If there are smutches on the instrument, clean them with cloth stained withalcohol.

This product is indoor instrument.

Power off when you finish your work. Pull off the connector plug whenthere’s long time no use of the instrument and cover it with a cloth orplastic paper to prevent from dust.

Pull the connector plug from the jack at once in the following case, andcontact the vendor:

There is some liquid flowing into the instrument;Drenched or fire burned;Abnormal operation: such as abnormal sound or smell;Instrument dropping or outer shell damaged;The function has obviously changed.

To assure the safe operation, please read carefully this manual beforeoperating.

CONTENTS

Chapter 1 Introduction------------------------------------------------------------------ -------1

Chapter 2 Specifications--------------------------------------------------------------- -------2

2.1 The normal operating condition----------------------------------------------------2

2.2 The basic parameters and performance-----------------------------------------2

Chapter 3 Preparations------------------------------------------------------------------------3

3.1 Structure description------------------------------------------------------------------3

3.2 Sample size requirements-----------------------------------------------------------4

3.3 Basic use for pedestal----------------------------------------------------------------4

Chapter 4 Software Setup--------------------------------------------------------------------6

4.1 Installation requirements-------------------------------------------------------------6

4.2 Installation preparations-------------------------------------------------------------6

4.3 Installation steps------------------------ -----------------------------------------------6

Chapter 5 Software operation--------------------------------------------------------------9

5.1 General introduction------------------------------------------------------------------9

5.2 Operation steps ------------------------------------------------------------------------11

5.3 Nucleic Acids ---------------------------------------------------------------------------12

5.4 Protein A280 ----------------------------------------------------------------------------15

5.5 Protein BCA-----------------------------------------------------------------------------17

5.6 Protein Lowry---------------------------------------------------------------------------21

5.7 Protein Bradford-----------------------------------------------------------------------24

5.8 UV-VIS Full-spectrum Scanning--------------------------------------------------28

Chapter 6 Failure Analysis and Troubleshooting-----------------------------------29

Operation Manual Chapter 1 Introduction

1

Chapter 1 Introduction

The SPECTRO-NANO is a spectrophotometer that measures 0.5ul-2ul samples with high

accuracy and reproducibility. Sample pedestals apply surface tension to make the

sample column, so that hold samples in the pedestal. During measurement, the light

goes through the sample column, and then passes to CCD detector by fiber optic

cable. In addition ，the SPECTRO-NANO has the capability to measure highly

concentrated samples without dilution(100X higher concentration than the samples

measured by a standard cuvette)

Operations Manual Chapter 2 Specification

2

Chapter 2 Specifications

2.1 The normal operating conditionAmbient temperature：5°C ∼ 35°CThe relative humidity：≤70%Power Supply：DC24V 2A

2.2 The basic parameters and performance

Model Parameter SPECTRO-NANO

Minimum Sample Size 0.5ul-2ul (2ul advised)

Path Length 0.2mm or 1mm

Light Source / Life Xenon flash lamp / >109 flashes

Detector Type 3864—element linear silicon CCD array

Wavelength Range 200－800nm

Wavelength Accuracy ±1 nm(TWHM at Hg 253.7nm

Spectral Resolution ≤3nm（FWHM@Hg 253.7nm）

Absorbance Precision 0.003Abs（1mm path length）

Absorbance Accuracy ±1％（7.332Abs, at 260nm wavelength）

Absorbance Range 0.04 - 90（at 260 wavelength, 10mm equivalent）

Detection Concentration Range 2ng/ul dsDNA ~ 4,500ng/ul dsDNA

Detection Time <10s

Input Voltage DC24V 2A

Power 20W

Dimension (W×D×H ) 200×250×166mm

Net Weight (kg) 2.6 kg

Software Compatibility Windows XP,Vista（32bit）,Win7（32bit/64bit）

Operations Manual Chapter 3 Preparations

4

3.2Sample size requirements

Although sample size is not critical, it is essential that the complete liquid column can beformed between the upper measurement pedestal and lower measurement pedestal tomake sure the precision of the measurement.It is best to use a precision pipettor (0-2ul) with precision tips to assure the precision ofthe sampling. If users are unsure about sample characteristics or pipettor accuracy, a 2ulsample is recommended.

3.3Basic use for the pedestal

3.3.1 With the upper pedestal open, pipette the sample (2ul) onto the lower pedestal.

3.3.2 Lower the sampling arm, the sample column is automatically drawn between the upperand lower measurement pedestals. Then the measurement initiates.

Upper measurementpedestal

Lower measurementpedestal

Sample column

Operations Manual Chapter 3 Preparations

5

3.3.3 When the measurement is complete, open the upper pedestal and wipe the samplefrom both the upper and lower pedestals using a soft laboratory wipe. Simple wipingprevents sample carryover in the pedestals.

Operations Manual Chapter 4 Software Setup

6

Chapter 4 Software Setup4.1 Installation requirement

Computer requirements：Microsoft Windows XP,Win7（32bit/64bit） or Vista（32bit）operating system1.5GHz or higher processor1GB or more of RAM（2GB for Vista operation system）

100MB of hard disk spaceOpen USB port（the instrument can only be connected with the computer via the USB port）

4.2Installation preparations

Before installing the software, please connect the instrument to computer as per photo below, thenturn on the power switch on instrument and the red light will on.

4.3Installation steps

4.3.1 Close all programs4.3.2 Insert CD to your driver and click “ ” to install.

Step1: Please select your language and click “OK”.

A

B


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Step2: Click “Next”

Step3: Choose Installation directory and click “Next”.

Step4：Make sure the installation directory is correct and then click “Install” to proceed, if you needto modify, please click “back”.


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Step5: After installation, there will be dialogue displayed as below, click “Next”.

Step6: Then there will be dialogue displayed as below, please click “Finish”.

Step7: After installation, there will be dialogue showed below, then please unplug and re plug inthe USB port once and click “OK”. In this way, the software installation is complete and you canclick the to run.

OK

Operations Manual Chapter 5 Software Operation

9

Chapter 5 Software Operation5.1 General IntroductionBefore sample measurement, connect the instrument with computer by using USB cable. Insertthe output end of power supply adapter (24V) into power connector in the back of instrument,and then open the power switch. In the forward side of instrument, power indication light is inred (green light is running indication light). When the instrument is not powered on, the plugneeds not to be unplugged. When the instrument is in the status of “stand by”, the power supplyis 5W, in this time, Xenon flash lamp is in the status of “close”.

5.1.1 Software characteristica) Application selection area

b) Measurement interface area

Save as text

Save as Picture


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There are two areas in SPECTRO-NANO software measurement interface. Working keys are in the left area, and displaying data are in the right area.

c) Task bar

The task bar include following options:[Home] – display the application main menu which can be operated by specified users.[My Data]- restore the sample data saved in the folder specified by the user.[Diagnostics]- test instrument if it is connected with computer properly.[Options]- update the machine parameters into user software.

Notice: When user changes the instrument (SPECTRO-NANO), before operation, please choose “Options” button, software will read new instrument’s data. After this, close software and restart software again, then operate instrument.

d) Function keys barWhen one function keys bar is clicked, in the top of left-side bar there will be 5 function keysas below.

Blank - Use dissolved sample buffer to make a blank. Before making a sample measurement,a blank must be measured.When blank measured，Absorbance testing value should be between 0.004-0.03 Abs.

Measure – Initiate the measurement for the samplesSave as text – The detail absorbance data of present measurement is stored in the txt table

which is specified by user.Save as picture –To store the current software interface imagePrint – Print a copy of current data in the default printer.


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e) Main menu barMain menu bar includes [File], [Tools], [Debug] and [Help].[File] includes the following options：Open- Restore the sample data saved in the folder specified by the user, equivalent to “My

Date” key.Save – Store result picture of present measurement.Print – Print a copy of current data in the default printer.Exit – Exit program

[Tools] includes the following options：Check connection –Test instrument if it is connected properly.Language - Select operation language.Factory set: Factory default setting

[Debug] includes the following options：DebugEnable: select if enter into Debug mode or not.LampEnable: select if open Xenon flash lamp.R_EliminateBlackI: select if retest dark currentSaveBlankData: save the blank data for this measurementOverlay spectra：The user can display 7 different colors (the

accumulation sequence is from red, orange,yellow, green, cyan, blue to purple) spectra in the data display area usingthis feature. After accumulate 7 different colors, they will be cleared.

[Help] includes the following options：Contents - Display electronic version of the manual.About- Display information about the software.

[Debug mode]

Integration time: the integration time for CCD sensor.iSampleNum: the average value for CCD sensor dataBoxcar width: the number of smooth filtering CCD pixelBoxcar Num: the number of smooth filtering for absorbance curve.

5.2 Operation stepsStep1: Twice click software icon, choose the software application (which the user is interest

in) in the right side bar.Step2: Use suitable buffer solution to build a blank. Load a 2ul blank sample onto the lower

pedestal and lower the upper pedestal into the 'down' position. Click 'Blank' button tomake a blank.

Step3: Wipe the sample from both the upper and lower pedestals using soft laboratory wipe.


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Input sample information (for instance: sample name) in the suitable location.Step4: Use 2ul sample and click “measurement” to make a measurement. Note: every time,

the each sample must be fresh.Step5: When the measurement is complete, a new laboratory wipe should be used to wipe

the pedestals. In this way, the users can do next measurement.

5.3 Nucleic Acids measurement5.3.1 General informationThe user can measure the concentration for nucleic acid by using SPECTRO-NANO.

If want to measure nucleic acids, select Nucleic Acid mode in the “main menu”The following equation is used to calculate the nucleic acids concentration:

c＝nucleic acids concentration (unit: ng/l)A＝absorbance value in AUε＝the wavelength-dependent extinction coefficient (unit: ng-cm/ul)b＝path length (unit: cm)The generally accepted extinction coefficients for nucleic acids are:Double-stranded DNA: 50ng-cm/ulSingle-stranded DNA: 33ng-cm/ulRNA: 40ng-cm/ulWhen selecting pedestal mode, SPECTRO-NANO Micro-spectrophotometer can measure high concentration nucleic acid sample without dilution from 1.0mm to 0.2mm short path length. The absorbance value of nucleic acid measurement is consistency of the reading value under 1cm path length.

The SPECTRO-NANO spectrophotometer can accurately measure double-stranded DNA samples up to 4500ng/ul without dilution. For each sample, software will automatically optimize the best path length to make measurement. When the optical intensity (after measurement sample extinction) is lower than 200(under 1cm path length), software will inform the customer to choose shorter path length to make sure the precision of the measurement. Unique screen is shown as below.


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The data shown in the absorbance spectrum image are consistency of the reading valueunder 10mm path length.

The right column of absorbance spectrum includes following information:Type－Used to select the type of nucleic acid being measured. The user can select

‘DNA-50’ for dsDNA, ‘RNA-40’ for RNA, ‘ssDNA-33’ for single-stranded DNA.Name－Input the sample name: when sample measurement, the sample name should be

input.ID--Display the serial number for the sample being measured. The user can open the

software to start recording.A260－Absorbance of the sample at 260nm with 10mm pathA280－Absorbance of the sample at 280nm with 10mm path260/280－Ratio of sample absorbance at 260nm and 280nm. The ratio of absorbance at

260 and 280nm is used to assess the purity of DNA and RNA. A ratio of ~1.8 isgenerally accepted as “pure” for DNA; a ratio of ~2.0 is generally accepted as“pure” for RNA. If the ratio is appreciably lower in either case, it may indicate thepresence of protein, phenol or other contaminants that absorb strongly at or near280nm.

260/230－ratio of sample absorbance at 260 and 230nm. This is a secondary measure ofnucleic acid purity. The 260/230 values of “pure” nucleic acid are often higherthan the respective 260/280 values. They are commonly in the range of 1.8-2.2. Ifthe ratio is appreciably lower, this may indicate the presence of co-purifiedcontaminants.

Baseline correction－If select the baseline correction, the default correction wavelength is340nm. The user can input different wavelength correction accordingto measurement requirements. In any measurement, the baseline isautomatically set to the absorbance value of selectable wavelength.All readings under wavelengths should be minus this value.

Note: The user may elect to turn off the baseline correction, which may result in the spectrabeing offset from the baseline.

5.3.2 Nucleic Acids concentration application steps1) In the main menu, select nucleic acid mode. It enters into the Nucleic Acidsmeasurement interface.

2) The users select the type of Nucleic Acid. The user can select 'DNA-50', 'RNA-40','ssDNA-33' and 'Other'. The default is DNA-50.

3) In the specific location, input sample name and item number.4) Create a blanking by using suitable solution. Blank solution，is normally solvent todissolve the targeting molecular and it need to be same with the samples in PH andionic strength. Load a 1-2ul blank sample onto the lower measurement pedestal andlower the sampling arm into the 'down' position. Click 'Blank' button to make a blank.When the correction is complete, the intensity curve will come out. Wipe the samplefrom both the upper and lower pedestals using soft laboratory wipe and measure thenext solution.

5) Load a 1-2ul ‘standby’ sample onto the lower pedestal and lower the sampling arm into


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the 'down' position. And then, click “Measure” key to do the measurement. When themeasurement is complete, the absorbance curve will come out. In the meantime, thedata for this measurement will show in the measurement result area. The defaultconcentration unit is ng/ul. The default standard wavelength is 340nm. If users want tochoose a new standard wavelength, they must enter into Debug mode. In the meantime,the measurement results will be added into the ‘measurement result list’ in the “List” of“TabPage”. If users want to see more detail absorbance intensity information, they cansee them in ‘ShowDetail’ in “List”. The absorbance intensity data can be stored in Excelfile by clicking ‘To Excel’. When click ‘My Data’ button to open the file, the measurementresult and absorbance curve will be restored in the software.

6) When the measurement is complete, a new laboratory wipe should be used to wipe thepedestals. In this way, the users can do next measurement. If measure the same lotsamples, the users do not need to re-blank. It is advisable to do the blanking every15min at least.

7) The software automatically adds the measurement results into the measurement resultslist in “List” of “TabPage”. It is best to click 'Save' button to save them in theuser-specified region before closing the software, since the data will disappear whenclosing the software.

Fig1. Nucleic Acids Measurement Result


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Fig2. Measurement Results List

5.4 Protein A2805.4.1 General information

Proteins, unlike nucleic acids, can exhibit considerable diversity. Protein A280 method isapplicable to purified proteins (includes Trp, Tyr residues or Cys-Cys disulfide) exhibitingabsorbance at 280nm. It does not require generation of a standard curve. The softwarecalculates the protein concentration directly after measure the absorbance value.The Protein A280 displays UV spectrum, measures the protein’s absorbance at 280nm andcalculate the concentration (mg/ml). Like the Nucleic Acids mode, it displays and records10mm equivalent data.

Measurement Concentration RangeThe SPECTRO-NANO Spectrophotometer will accurately measure protein samples up to 90mg/ml BSA) without dilution. When the optical intensity (after measurement sample extinction) is lower than 200(under 10mm path length), software will inform the customer to choose shorter path length to make sure the precision of the measurement. Unique screen is shown as below.

The hydrophobic between the water molecules is the main factor of surface tension. Ingeneral, the presence solute of liquids (（ including protein, DNA, RNA, salt ion, detergentmolecule) can significantly reduce surface tension. Although, for most samples, a 1ul samplesize is enough, a 2ul sample size is recommended for protein measurements that the liquidcolumn be formed.

Screen display:


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The data shown in the absorbance spectrum image are consistency of the reading value at10mm path length.

The right column of absorbance spectrum includes following information:Type－Used to select the type of nucleic acid being measured. The user can select ‘A280’

for A280, ‘BSA’ for BSA, ‘lgG’ for lgG, and 'Lysozyme' for Lysozyme.Name－Input the sample name: when sample measurement, the sample name should be

input.ID--Display the serial number for the sample being measured. The user can open the

software to start recording.A260－Absorbance of the sample at 260nm with 10mm pathA280－Absorbance of the sample at 280nm with 10mm path260/280－Ratio of sample absorbance at 260nm and 280nm.260/230－ratio of sample absorbance at 260 and 230nm.Baseline correction－If select the baseline correction, the default correction wavelength is

340nm. Users can input different wavelength correction according tomeasurement requirements. In any measurement, the baseline isreadings under wavelengths should be minus this value.

Note: The user may elect to turn off the baseline correction, which may result in the spectrabeing offset from the baseline.

5.4.2 Protein A280 concentration application steps1) In the main menu, select Protein A280 mode. It enters into the protein measurementinterface.

2) The users select the type of Protein. The user can select 'A280', 'BSA', 'lgG' and'Lysozyme'. The default is A280.


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3) In the specific location, input sample name and item number.4) Create a blanking by using suitable solution. Blank solution，is normally solvent to dissolvethe targeting molecular and it need to be same with the samples in PH and ionic strength.Load a 1-2ul blank sample onto the lower measurement pedestal and lower the sampling arminto the 'down' position. Click 'Blank' button to make a blank. When the correction is complete,the intensity curve will come out. Wipe the sample from both the upper and lower pedestalsusing soft laboratory wipe and measure the next solution.

5) Load a 1-2ul ‘standby’ sample onto the lower pedestal and lower the sampling arm into the'down' position. And then, click “Measure” key to do the measurement. When themeasurement is complete, the absorbance curve will come out. In the meantime, the data forthis measurement will show in the measurement result area. The default concentration unit ismg/ml. The default standard wavelength is 340nm. If users want to choose a new standardwavelength, they enter into Debug mode. In the meantime, the measurement results will beadded into the ‘measurement result list’ in the “List” of “TabPage”. If users want to see moredetail absorbance intensity information, they can see them in ‘ShowDetail’ in “List”. Theabsorbance intensity data can be stored in Excel file by clicking ‘To Excel’. When click ‘MyData’ button to open the file, the measurement result and absorbance curve will be restoredin the software.6) When the measurement is complete, a new laboratory wipe should be used to wipe thepedestals. In this way, the users can do next measurement. If measure the same lotsamples, the users do not need to re-blank. It is advisable to do the blanking every 15minat least.

7) The software automatically adds the measurement results into the measurement results listin “List”. It is best to click 'Save' button to save them in the user-specified region beforeclosing the software, since the data will disappear when closing the software.

5.5 Protein BCA5.5.1 General information

The BCA Protein Assay is an alternative method for determining protein concentration. It isoften used for more dilute protein solutions and/or in the presence of components that alsohave significant UV absorbance. Unlike the Protein A280 method, the BCA Assay requires astandard curve to be generated each time it is run, before unknown proteins can bemeasured. BCA Assay is testing Cu+1 ion, under alkaline environment, Cu＋2 ion will bereconditioned to CU+1 by protein. Two Biquinoline 2-dicarboxylic acids BCA molecular andone Cu+1 ion will form purple chelate in the presence of protein. Cu-BCA chelate ismeasured at its wavelength maximum of 562nm and normalized at 750nm.

5.5.2 BCA Assay Measurement RangeCommercial BCA kit procedures for two different protein measurement range:a) A regular assay — using a 20:1 reagent/sample volume ratio. This kit measurement

range is from 0.20mg/ml to 8.0mg/ml (BSA). When pedestal measuring, we suggestusing sample volume of 4ul and 80ul BCA reagent.b) A mini assay — using a 1:1 reagent/sample volume ratio. To prepare enough samplevolume for pedestal measurement, range from 0.01mg/ml to 0.20mg/ml. We suggest using


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10ul samples and 10ul BCA reagent in PCR tubes.In addition to the kit reagents, protein standards (BSA) for generating a standard curve areprovided for the BCA method by the manufacturer. Make sure that all measurements use thesame incubation times and temperature.

Note: If the Ambient Temperature is higher than 60℃, please double the sample size toavoid the volatilization.

5.5.3 Unique Screen FeaturesBCA unique screen is as below:

The spectrum image shows present sample data. The measurement results by using pedestalare consistency of the reading value at 10mm path length.

5.5.4 BCA Standard CurveA standard curve is required every time the BCA assay is run.


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5.5.5 Standard Curve Set-upA simplest standard curve includes three points, including multi-point standard curve and areference (BCA reagent only – no protein). Up to 7 standard points can be measured. Thereis no set order in which standard points must be run.Note: standard points can only be added to standard curve before making measurements

for the samples. After adding, the user cannot add or delete any standard point intothe standard curve during making measurement. The concentration range of standardpoints should cover all standby sample concentration.

5.5.6 Making BCA Measurements1) Choose BCA in the main menu.

2) Enter the BCA interface.

Add the reference

and standard sample

concentration value

Click it to read the

absorbance of each

line

Choose BCA


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3) Add the banking sample, and then click “Bank” to make a bank.4) Add reference and standard sample concentration value. (The input value must be thevalue from 0 to 2000, to three decimal places.)

5) Add reference and corresponding samples, click each-line reference sample and standardsample by using the right mouse button. Then read the corresponding absorbance. (Theuser can read 4-times absorbance values at most to make the average value)

6) Click “Input” to draw a standard curve.7) Add “standby sample”, and click “Measure” to make the measurement.

8) Finish the measurement.

Click “Input” to draw

a standard curve

Standard curve

Result curve

Measurement record


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5.6 Protein Lowry5.6.1 General information

The Lowry Protein Assay is an alternative method for determining protein concentrationbased on the widely used and cited Lowry procedure for protein quantitation. Like otherAssays, the Lowry Assay requires standard curve generation each time it is run. The Lowryprocedure involves reaction of protein with cupric sulfate in alkaline solution, resulting information of tetradentate copper protein complexes. The Folin－Ciocalteu Reagent iseffectively reduced in proportion to the chelated copper-complexes resulting in awater-soluble blue product that is measured at 650nm and normalized at 405nm. Thereagents utilizing in the assay, are available in kit form from numerous manufacturers.

5.6.2 Measurement Concentration RangeTo accurately prepare standards, a sample volume of 20ul and 100ul of Lowry reagent is recommended. On the SPECTRO-NANO Spectrophotometer, the Lowry assay can run from 0.20mg/ml to 4.0mg/ml. Follow the manufacturer’s protocol for the assay. Make sure that all measurements use the same incubation time and temperature. In addition to the kit reagents, protein standards (BSA) for generating a standard curve are provided for the Lowry Protein method by the manufacturer. Since the SPECTRO-NANO Micro-Spectrophotometer can measure higher protein concentrations, you may need to supply your own protein standards at higher concentrations than provided by the manufacturer. A 2ul sample size is recommended for protein measurements.

5.6.3 Unique Screen FeaturesLowry unique screen is as below:


5.6.4 Lowry Standard CurveA standard curve is required every time the Lowry assay is run.

SPECTRO-NANO Operations Manual Chapter 5 Software Operation

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5.6.5 Standard Curve Set-upA simplest standard curve includes three points, including multi-point standard curve and areference (Lowry reagent only – no protein). Up to 7 standards can be measured. There isno set order in which standards must be run.Note: standards can only be added to standard curve before making measurements for the

samples. After adding, the user cannot add or delete any standard point into thestandard curve during making measurement. The concentration range of standardpoints should cover all standby sample concentration.

5.6.6 Making Lowry Measurements1) Choose Lowry in the main menu.

Choose Lowry


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2) Enter the Lowry interface.




Add the reference

and standard sample

concentration value


absorbance of each

line

Click “Input” to

draw a standard

curve

Standard curve


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5.7 Protein Bradford5.7.1 General information

The Bradford Assay is an alternative method commonly utilized for determining proteinconcentration. It is often used for more dilute protein solutions where lower detectionsensitivity is needed. Like the BCA and Lowry Assays, the Bradford Assay requires astandard curve. The bradford uses the protein-induced absorbance shift of Coomassie Bluedye to 595 nm as a measurement of protein concentration. The bound protein-dye complexis measured at 595 nm and normalized at 750 nm. Correspondent kits are available fromnumerous manufacturers.

5.7.2 Measurement Concentration RangeCommercial Bradford Protein kit manufacturers typically outline procedures for two differentconcentration ranges:a) A regular assay — using a 50:1 reagent/sample volume ratio. This kit measurement

range is from 0.10mg/ to 8.0mg/ml（BSA）. The best linearity is in the 0.01-1mg/ml. Whenpedestal measuring, we suggest using sample volume of 4ul and 200ul Bradford reagent.b) A mini assay — using a 1:1 reagent/sample volume ratio. To prepare enough samplevolume for pedestal measurement, range from 15ug/ml to 125ug/ml. We suggest using 10ul samples and 10ul BCA reagent in PCR tubes.In addition to the kit reagents, protein standards (BSA) for generating a standard curve are provided for the Bradford method by the manufacturer. Make sure that all measurements use the same incubation times and temperature.Note: If the Ambient Temperature is higher than 60℃, please double the sample size to avoid the volatilization.Since the SPECTRO-NANO Micro-Spectrophotometer can measure higher protein concentrations, you may need to supply your own protein standards at higher concentrations than provided by the manufacturer.

Result curve

Measurement

records


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5.7.3 Unique Screen FeaturesBradford unique screen is as below:


5.7.4 Bradford Standard CurveA standard curve is required every time the Bradford assay is run.


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5.7.5 Standard Curve Set-upA simplest standard curve includes three points, including multi-point standard curve and areference (Bradford reagent only – no protein). Up to 7 standards can be measured. Thereis no set order in which standards must be run.Note: standards can only be added to standard curve before making measurements for the

samples. After adding, the user cannot add or delete any standard point into thestandard curve during making measurement. The concentration range of standardpoints should cover all standby sample concentration.

5.7.6 Making Bradford Measurements1) Choose Bradford in the main menu.

2) Enter the Lowry interface.

Choose Bradford

Add the reference

and standard sample


absorbance of each

line


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Click “Input” to

draw a standard

curve

Standard curve

Result curve

Measurement

records


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5.8 UV-VIS Full-spectrum Scanning5.8.1 General informationUV-VIS module allows the SPECTRO-NANO Spectrophotometer to function as a conventional

spectrophotometer. Sample absorbance is displayed on the screen from 200nm to 800nm.

5.8.2 Measurement Concentration RangeSamples with high absorbance (up to 90A equivalent at 10mm path) can be measureddirectly.

5.8.3 Unique Screen Features1) Choose “UvVis” in the main menu.

2) Measurement result interface

Blank

Operations Manual Chapter 6 Failure Analysis and Troubleshooting

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Chapter 6 Failure Analysis and Troubleshooting

Failure analysis and processing procedures

No. Fault phenomenon Cause analysis Recovery processing

1The indicator light is notbright the instrument ispowered on.

No power Check the connection of power

Broken switch Exchange the switch

Broken Power Line Contact to the seller

Others Contact to the seller

2 The instrument can not beconnected with computer

The instrument is notpowered on Power on the instrument

Poor USB cable Change the USB cable

Documents

SPECTRO-NANO - MRCLAB · Operation Manual SPECTRO-NANO ... Step5: After installation, ... Language - Select operation language. Factory set: Factory default setting