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Agilent 2200 TapeStation - User Manual

1. Agilent Technologies will not sell these parts separately Ifan IKA MS3 vortexer is not available please ensure thorough manual vortex mixing 10 seconds on maximum speed For successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape For best sizing precision and accuracy run the appropriate ladder with the samples If 16 samples need to be analysed in parallel you may choose to insert a software ladder in the 2200 TapeStation analysis software However sizing may not be as accurate with a software ladder No Software ladder is available for the Genomic DNA application 58 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation D1000 ScreenTape Assay Parts required Parts required 1 uL DNA or Ladder Prepare TapeStation D1000 p n Description 5067 5582 D1000 ScreenTape 1 Launch the Agilent 2200 TapeStation software 2 Load D1000 ScreenTape and loading tips into the 2200 TapeStation Sample Preparation D1000 ScreenTape Assay p n Description 5067 5583 D1000 Reagents D1000 Ladder D1000 Sample Buffer Allow reagents to equilibrate at room temperature for 30 min 2 Vortex mix before use 3 If running ladder add 3 uL D1000 Sample Buffer 9 to 1 uL D1000 Ladder 4 Add 3 uL D1000 Sample Buffer 9 to 1 uL DNA sample 5 Vortex usin
2. Jaami O O Agilent Technologies Sample Descriptions Sample Well Description l 12 ODOQDOOQOOOO 0100101010101010 ofolofolofololo Totelelelelelelelelelere D1000 ScreenTape ID 00 S029 010109 99 138699 Expires 29 Oct 2013 Ready Figure 6 Controller image 96 well plate selection In the 96 well plate sample selection screen the panel labeled LDR is not available for selection All information entered in the control software will appear in the analysed results Agilent 2200 TapeStation User Manual 43 4 Using the 2200 TapeStation System Start the TapeStation Run 1 Click the start button This will produce a Save As window As a default the file name starts with the date in reverse order and a run counter When run continuously the save function auto increments the counter part of the file name yore Gp 2200 TapaSusion Dats File pone ot 10 14 09 Save or pe Fi T apeStaton O ata Fe Fi 2 Type in the name that you wish the analysis to be saved as Do not include a full stop in file names Final Check ScreenTapeController TapeStation Controller 1 Please ensure tips and samples are loaded close the lid and click OK to continue 1 Lift the lid of the TapeStation 2 Ensure that there are fresh tips in the tip holder and that all the samples have been correctly loaded with lids removed and correspond to the sample selection on the screen 3 Close the lid Lift
3. The presence of small bubbles within the buffer chamber of the ScreenTape is normal These bubbles often occur at the gel buffer interface and need to be displaced prior to running Failure to remove bubbles from the gel buffer interface is detrimental to the performance of the ScreenTape 6 Insert the ScreenTape into the TapeStation with the label towards the front of the instrument and the barcode facing right Protect the individual gel lanes within the ScreenTape from excessive force Do not bend or flex ScreenTape and store in the provided packaging at the recommended temperature when not in use TapeStation instrument will not recognize the screen tape if inserted incorrectly The TapeStation will automatically recognise the sample plate type and ScreenTape and load the required parameters Agilent 2200 TapeStation User Manual CAUTION Using the 2200 TapeStation System 4 7 Prepare samples according to type as detailed in How to prepare your samples on page 48 or the appropriate Quick Guide 8 Place samples into the sample block inside the TapeStation Damage to the 2200 TapeStation and impact on performance gt Ensure the lids have been removed from the sample tubes 9 Select the tubes or wells you wish to run by clicking and dragging the mouse over the sample locations Selected wells will change colour from white Selected lanes on the controller ScreenTape image will change colour Lanes
4. Introduction to the 2200 TapeStation System Information pertaining to sample and ScreenTape requirements can be found in ScreenTape Architecture Operating Procedure ScreenTape Architecture Barcode Buffer chamber Electrodes Gel ScreenTape product details Front View ScreenTape M details t 2 4 i C M E H 4 15 H A a 5 Hn LE 3 ea H Buffer Chamber gt lC t I EL LN H LE M ea H 4 la ve HA ay H Electrodes 3 7 la I la EM r lA 1 d amp gt 1 4 HM Figure 1 Example P200 ScreenTape Agilent 2200 TapeStation User Manual The unique barcode tracks lane usage within individual ScreenTape and allows traceability of results The buffer chamber is located at the top of the channel and contains optimised buffers for the effective separation of nucleic acid fragments or proteins The integrated electrodes apply a current across the ScreenTape and eliminate the need for any additional electrophoresis equipment The gel contained within ScreenTape has been developed specifically to resolve nucleic acids or proteins The information is unique to each consumable item This includes ScreenTape type product expiry date and a unique serial number Back View 1 1 Introduction to the 2200 TapeStation System Agilent 2200 TapeStation Components Lid The 2200 TapeStation lid must be closed each time the inst
5. 64 Using the 2200 TapeStation System Mixing recommendations TapeStation instruments are supplied with an optional IKA MS3 vortexer which includes a 96 well plate adaptor suitable for both 96 well PCR plates and 8 way strips This vortexer is recommended for use with the following applications D1000 ScreenTape Assay High Sensitivity D1000 ScreenTape Assay RNA ScreenTape Assay High Sensitivity RNA ScreenTape Assay tis recommended that all current TapeStation users purchase this instrument direct from IKA for use with these assays Please quote part numbers 5067 5700 and 4674100 when ordering Agilent Technologies will not sell these parts separately Ifan IKA MS3 vortexer is not available please ensure thorough manual vortex mixing 10 seconds on maximum speed Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation RNA ScreenTape Assay Parts required Parts required Prepare TapeStation p n Description 5067 5576 RNA ScreenTape 1 Launch the Agilent 2200 TapeStation software 2 Load RNA ScreenTape and loading tips into the 2200 TapeStation 3 Select RNA Protocol Eukaryotic RNA or Prokaryotic RNA Sample Preparation RNA ScreenTape Assay p n Description 5067 5577 RNA ScreenTape Sample Buffer 5067 5578 RNA ScreenTape Ladder 1 Allow reagents to equilibrate at room temperature for 30 min 2 Vortex mix before use 3 Thaw total RNA samples on ice 4 Add 5 uL Sample Buf
6. User Manual p Agilent Technologies Notices Agilent Technologies Inc 2011 2012 2013 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 62964 90002 Rev B Edition 10 2013 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn This product may be used as a com ponent of an in vitro diagnostic sys tem if the system is registered with the appropriate authorities and com plies with the relevant regulations Otherwise it is intended only for gen eral laboratory use Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connection with the furnishing use or perfor mance of this document or of any information contained herein Should Agilent and the
7. indicating the software did not pass the windows logo testing click Continue Anyway A window appears indicating that the hardware has been successfully installed The TapeStation system will function As there is more than one driver that can be detected and installed you may need to follow these steps more than once You may need to follow these steps if you change the USB port on the laptop for the TapeStation connector cable Agilent 2200 TapeStation User Manual 29 3 Installing the System Installing the System 30 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual 4 Using the 2200 TapeStation System Intended Use of the 2200 TapeStation System 33 Performance Limitations of Use 34 Additional Components Required by the User 35 Operating Procedure 36 Add Experiment Notes 41 Describe Samples 42 Start the TapeStation Run 44 Final Check 44 Running System 45 Abort the TapeStation Run 45 Complete TapeStation Run 45 Empty Tip Buckets 46 How to Use the Agilent TapeStation Software 46 Shutdown and Restarting Procedure 47 How to prepare yoursamples 48 Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation 50 Quantification 50 Sizing 53 GenomicDNA 54 Molarity 56 DNA Sample Preparation 57 Sample Preparation D1000 ScreenTape Assay 59 Sample Preparation High Sensitivity D1000 ScreenTape Assay 60 Sample Preparation Genomic DNA ScreenTape Assay 61 RNA Sample Preparation 63 Sam
8. 82 WEEE directive 82 Agilent 2200 TapeStation User Manual www agilent com In This Book The manual describes the following Introduction to the system Site requirements and specifications Installation Using the system Maintenance Product notices O Agilent Technologies 2011 2012 2013 Printed in Germany 10 2013 G2964 90002 Rev B RES Agilent Technologies
9. Agilent 2200 TapeStation User Manual 23 3 Installing the System Contents of the ScreenTape System The Agilent 2200 TapeStation Table1 The Agilent 2200 TapeStation System G2964AA G2965AA Product Volume Properties Agilent 2200 1x Instrument for loading electrophoresing imaging and analysing TapeStation 2200 TapeStation System G2964AA or e 2200 TapeStation Nucleic Acid System G2965AA TapeStation Software 1x CD The software is required to drive the 2200 TapeStation and visualise the Setup Disc ScreenTape analysis Laptop 1x Laptop Instrument Control Laptop USB Cables Power 1 x USB cable 1 x USB cable to connect the laptop to the TapeStation supply units 2 x power cords 1 x Power supply unit for the laptop 1 x Power supply unit for the TapeStation Sample Block 1 x02 mL strip A removable sample block for the correct loading of samples within the and TapeStation 1 x96 well plate Tip Holder 2x A removable cartridge for pipette tips placed in the TapeStation TapeStation loading tips TapeStation compatible 0 2 mL tube strips and lids 96 well plates 96 well plate foil seal Loading tip transfer tool optional IKA Vortexer and associated PCR plate optional Guides 1x384 tips 1x box of 120 tubes and caps pack of 10 pack of 100 1x 1x Pipette tips to use in the 2200 TapeStation Tube strips for placing samples mixed with loading buffer into the 2200 TapeStation Vortexer for mixing samples and
10. C d After heating remove condensation from the lids or foil cover of the tubes by centrifugation P200 pH buffer clear is supplied to allow the user to dilute samples to alleviate issues with staining efficiency caused by low pH The use of P200 pH Buffer resolves these issues in most circumstances For further information on buffer compatibility contact your Agilent Technologies representative 3 Denaturate sample a Choose which sample buffer is required P200 Reducing Sample Buffer O or P200 Non reducing Sample Buffer It is recommended that P200 Reducing Sample Buffer is used for the denaturation of P200 Ladder b Add 4 uL of the relevant P200 sample buffer to the stained sample and replace the lids or foil cover c Mix then heat at 75 C for 5 min d Remove condensation from the lids or foil cover of the tubes by centrifugation 4 Add 2 uL of P200 Marker 8 to each sample and to the P200 ladder Agilent 2200 TapeStation User Manual 7 4 Using the 2200 TapeStation System Protein Sample Preparation 5 Mix the solution well and centrifuge to ensure that the sample is at the bottom of the tube ready for analysis on the TapeStation NOTE P200 Marker is formulated with a high percentage of glycerol Due to the high density of this reagent the user must ensure that the samples are adequately mixed prior to analysis on the TapeStation Failure to do so may result in unsatisfactory analysis results 6 Loa
11. ScreenTape Specifications 14 This chapter provides information on specifications de Agilent Technologies 2 12 Specifications Technical Specifications 2200 TapeStation Input voltage Power consumption Current Interface Instrument Housing Dimensions Weight Power Supply Input voltage Input frequency Phase Current 12V DC 40 W VA 3A USB cable PC comms UL94 VO rated flame retardant cast polyurethane 400 x 310 x 310 mm 12 5 kg 100 240 V AC 50 60 Hz 1 0 45 1 1A Environmental condition Optimal operating temperature Instrument operating temperature 23 C 73 4 F 12 37 C 53 6 98 6 F for D1000 ScreenTape 17 37 C 62 6 98 6 F for High Sensitivity D1000 ScreenTape 15 30 C 59 0 86 0 F for Genomic DNA ScreenTape 14 30 C 57 2 86 0 F for RNA and High Sensitivity RNA ScreenTape 10 33 C 50 0 91 4 F for P200 ScreenTape 1 prolonged use Instrument operating temperature may be higher than ambient lab temperature especially after Agilent 2200 TapeStation User Manual Specifications 2 If the instrument is out of the recommended temperature range for the ScreenTape inserted the following error message will appear in the controller software 2200 TapeStation Controller 01 04 pem Ladder Needle Change Transport About pit Agilent Technologies if Sample De
12. Specifications High Sensitivity D1000 ScreenTape and Reagents Sizing Range Typical Resolution Sensitivity Sizing Precision Sizing Accuracy Quantitative Precision Quantitative Accuracy Quantitative Range Physical Specifications Analysis Time Samples per consumable Sample volume required Kit stability Kit size 35 1000 bp 35 300 bp 15 300 1000 bp 10 5 pg uL 5 CV 10 15 CV 20 10 1000 pg uL 16 samples 20 min 96 samples 100 min 16 2 uL 4 months 112 samples 1 signal to noise gt 3 single peak 2 Sizing Accuracy for software ladder 20 3 Measured against 2100 Bioanalyzer Agilent 2200 TapeStation User Manual 15 2 16 Specifications Specification Genomic DNA ScreenTape Assay Analytical Specification Genomic DNA ScreenTape and Reagents Sizing Range Sensitivity Sizing Precision Sizing Accuracy Quantitative Precision Quantitative Accuracy Linear Concentration Range Carry Over Physical Specification Analysis Time Samples per consumable Sample Volume Required Shelf Life Box Kit size 200 bp to gt 60000 bp 0 5 ng pL 200 15000 bp 15 6 CV 200 15000 bp 10 15 96 CV 20 6 10 100 ng uL N A 16 samples 25 min 96 samples 150 min 16 Tul 4 months 112 samples box 1 Determined using the Genomic DNA Ladder as sample Average result from various genomic DNA sample type
13. are RNase free gt Handle RNA samples with care gt Wear gloves at all times gt Thaw RNA samples on ice gt Vortex and centrifuge all samples before use gt Store RNA samples on ice throughout the ScreenTape analysis procedure Itis important to place the samples on ice directly after the denaturation step as this aids complete and stable denaturation of the RNA To ensure optimal performance of the RNA ScreenTape platform samples should be analysed using the 2200 TapeStation within 3 h of the denaturation step when left on the 2200 TapeStation system Beyond 3 h denatured samples should be stored on ice or in a suitable freezable sample block For best results ensure that all reagents are allowed to equilibrate to room temperature prior to use When pipetting Sample Buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to the viscosity of Sample Buffers When pipetting small volumes ensure that no sample remains within the tip When adding Sample Buffer to sample please ensure thate they are mixed correctly To achieve this gently mix several times with additional pipetting then cap the tubes vortex mix using IKA vortexer and adaptor at 2000 rpm for 1 min Briefly centrifuge to collect the contents at the base of the tubes Improper mixing can lead to quantification errors Agilent 2200 TapeStation User Manual 63 4
14. reagents Site Safety guide and Quick Guides 62964AA Protein DNA and RNA G2965AA DNA and RNA 24 Agilent 2200 TapeStation User Manual ScreenTape Products Installing the System 3 Kit Components High Sensitivity D1000 ScreenTape Assay Part Number Name Color Amount 5067 5584 High Sensitivity D1000 ScreenTape 7 ScreenTape 5067 5585 High Sensitivity D1000 Regents 2 vials High Sensitivity D1000 Ladder o 20 uL High Sensitivity D1000 Sample Buffer O 300 pL 5067 5587 High Sensitivity D1000 Ladder 1 vial o 20 uL Kit Components D1000 ScreenTape Assay Part Number Name Color Amount 5067 5582 D1000 ScreenTape 7 ScreenTape 5067 5583 D1000 Reagents 2 vials D1000 Ladder o 10 pL D1000 Sample Buffer O 400 pL 5067 5586 D1000 Ladder 1 vial o 10 uL Kit Components Genomic DNA ScreenTape Assay Part Number Name Color Amount 5067 5365 Genomic DNA ScreenTape 7 ScreenTape 5067 5366 Genomic DNA Reagents 2 vials Genomic DNA Ladder o 75 uL Genomic DNA Sample Buffer O 1350 uL Agilent 2200 TapeStation User Manual 25 3 Installing the System Kit Components High Sensitivity RNA ScreenTape Assay Part Number Name Color Amount 5067 5579 High Sensitivity RNA ScreenTape 7 ScreenTape 5067 5580 High Sensitivity RNA ScreenTape Sample 1 vial Buffer e 250 uL 5067 5581 High Sensitivity RNA ScreenTape Ladder 1 vial o 10 uL Kit Components RNA ScreenTape Assay Part Number Name Color Amou
15. Additional Consumables required for the 2200 TapeStation instrument Loading tips 5067 5152 Loading tips 5067 5153 Optical Tube 8x Strip Box of 120 401428 and Optical Cap 8x Strip Box of 120 401425 or well Sample Plates Pack of 10 plates 5067 5150 and well Plate Foil Seal Pack of 100 foils 5067 5154 Vortex mixer See note below Additional Material Required Not Supplied Volumetric pipette Centrifuge Heating block or PCR machine Mixing recommendations TapeStation instruments are supplied with an optional IKA MS3 vortexer which includes a 96 well plate adaptor suitable for both 96 well PCR plates and 8 way strips This vortexer is recommended for use with the following applications D1000 ScreenTape Assay High Sensitivity D1000 ScreenTape Assay RNA ScreenTape Assay High Sensitivity RNA ScreenTape Assay It is recommended that all current TapeStation users purchase this instrument direct from IKA for use with these assays Please quote part numbers 5067 5700 and 4674100 when ordering Agilent Technologies will not sell these parts separately If an IKA MS3 vortexer is not available please ensure thorough manual vortex mixing 10 seconds on maximum speed Agilent 2200 TapeStation User Manual 35 4 Using the 2200 TapeStation System Operating Procedure 1 Double click the 2200 TapeStation controller icon on the desktop and follow the instructions on the screen Always ensure you are usi
16. Buffer 9 to 2 uL RNA sample or diluted RNA ScreenTape Ladder C For best results use the reverse pipetting technique Freeze any unused Ladder at 20 C 6 Vortex using IKA vortexer and adaptor at 2000 rpm for 1 min 7 Spin down to position the sample at the bottom of the tube Agilent 2200 TapeStation User Manual 67 4 Using the 2200 TapeStation System RNA Sample Preparation 8 Sample denaturation a Heat samples to 72 C for 3 min b Place samples on ice for 2 min c Centrifuge to collect samples in the base of the tubes 2 uL RNA sample or 1 uL HS RNA HS RNA Sample Buffer Heat at 72 C Ladder for 3 minutes pi M D dE um Sensitivity PI i RNA Vortex 1 min vue p Sfin dowi for 2 minutes 68 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Protein Sample Preparation When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to viscosity of Sample Buffers When pipetting small volumes ensure that no sample remains within the tip When adding sample buffer to sample please ensure that they are mixed correctly by following assay instructions mproper mixing can lead to quantification errors Once mixed briefly centrifuge to collect the contents at the base of tubes Forbestresults ensure that all reagents are allowed to equilibrate to room temperature for 30 minutes prio
17. ange is recommended 76 Agilent 2200 TapeStation User Manual Maintenance 5 Parts required p n Description 1 G2960 60062 Needle cartridge for use in single pump systems For use with product numbers ST007 ST008 and ST009 OR 1 G2960 60063 Needle cartridge for use in dual pump systems For use with product numbers ST017 ST019 ST010 G2960A G2961A G2964AA G2965AA and G2966AA New needles cartridges can be ordered at any time from Agilent Technologies by contacting your local sales agent For details on correct needle cartridge for your TapeStation model refer to Table 3 on page 75 Change the needle cartridge 1 Remove the sample plate and tip holder 2 Remove the foil tab from the top of the needle cartridge Care must be taken to keep the needle cartridge level after removing the foil tab 3 Insert the needle cartridge into the tip holder space using the label for orientation The cartridge should be placed so that the label faces to the right and the printed arrow points to the front of the TapeStation Needle cartridge Rear TapeStation Front 4 Close the lid 5 Go to Needle Change on the Controller software toolbar and select Run Agilent 2200 TapeStation User Manual 71 5 Maintenance Changing the Needle 78 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual 6 Appendix Limited Use Label License 80 Sound Emission 81 The Waste Electrical and Electronic E
18. d sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section 2uL Protein 2yL 2yL sample Or Stain P200 P200 Solution markers Ladder Vortex Vortex Heat at 75 C Heat at 75 C for 7 minutes for 5 minutes 72 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual e e 9 e Maintenance e e e e General Information 74 Changing the Needle 75 This chapter describes the maintenance of the TapeStation system ee Agilent Technologies 73 5 Maintenance General Information 74 Annual Preventative Maintenance PM is essential for the TapeStation as it has many moving parts This PM should be arranged with your local Agilent representative and consists of Fan Filter replacement Needle replacement Electrophoresis probe replacement Internal instrument inspection for wear foreign objects and general clean inside and out In addition to the above the engineer will check that the instrument is functional by running a full ScreenTape For customers with exceptionally high usage the needle replacement procedure as detailed in the section below can also be performed between annual PM services Agilent 2200 TapeStation User Manual Changing the Needle Table3 Overview TapeStation Configuration Needle Cartridge Maintenance 5 It is important to know which TapeStation system you have before changing the needle s in order
19. e of the product or its components to provide a service information or data 3 use of the product or its components for therapeutic diagnostic or prophylactic purposes or 4 resale of the product or its components whether or not such product or its components are resold for use in research For information on purchasing a license to this product for purposes other than research contact Molecular Probes Inc Business Development 29851 Willow Creek Road Eugene OR 97402 USA Tel 541 465 8300 Fax 541 335 0354 Agilent 2200 TapeStation User Manual Appendix 6 Sound Emission Manufacturer s Declaration This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991 This product has a sound pressure emission at the operator position lt 70 dB Sound Pressure Lp lt 70 dB A At Operator Position Normal Operation According to ISO 7779 1988 EN 27779 1991 Type Test Agilent 2200 TapeStation User Manual 81 6 Appendix The Waste Electrical and Electronic Equipment Directive Abstract The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC adopted by EU Commission on 13 February 2003 is introducing producer responsibility on all electric and electronic appliances starting with 13 August 2005 This product complies with the WEEE Directive 2002 96 EC marking requirements The affixed label indicates that you must not di
20. e prepared stain solution is best used on the day of formulation however it can be stored for up to one week below 20 C For normal applications 2 uL of formulated stain solution is required for 2 uL of sample For 16 samples 8 uL of 5X Stain would be diluted with 32 uL of Stain Buffer The resultant stain solution should be thoroughly mixed before use For certain applications particularly with high protein concentrations higher concentrations of stain can be used in combination with altered ratios of stain to sample 8 uL P200 5x A Labeling Solution dye P200 For 16 samples prepare 40 pL stain solution Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 2 Stain protein sample or ladder The P200 ladder C should be processed through the P200 sample preparation procedure in the same manner as your samples In Ladder mode selected in the ladder options in the controller software P200 ladder is automatically selected as the first sample in the TapeStation controller The user can also select to run no ladder and then to insert a software ladder in the 2200 TapeStation Analysis software However sizing may not be as accurate with a software ladder a Place 2 uL of P200 stain solution prepared above into a PCR tube strip or 96 well plate b Pipette 2 uL of the protein sample or ladder into the tube mix and attach the lids or foil cover to prevent evaporation c Heat for 7 min at 75
21. elections 40 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Add Experiment Notes 1 If required notes can be manually entered into the software before the instrument is started Ladder NeedleChange Transport About Agilent Technologies Sample Descriptions Sample Well Description om Oo O O oO Oo OQ O O O O O OQ Oo OQ O O D E F G H D1000 ScreenTape Q Agilent Technologies ID 00 5029 010109 99 138699 Expires 29 Oct 2013 5 Ready Agilent 2200 TapeStation User Manual 41 4 Using the 2200 TapeStation System Describe Samples 1 Sample descriptions can be manually entered into the software before the instrument is started and whilst the TapeStation is operating before analysis software is launched OR Sample data can be copied and pasted from an Excel table The entered Sample descriptions data can be copied to clipboard by using the Copy to Clipboard link in the top right hand corner of the Sample Description table 42 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Ladder Needle Change Transport About Agilent Technologies Germent 135 om Sample Descriptions t a Sample Well Description o 0 0 00 0010 jejejejejejeje o D1000 ScreenTape ID 00 S029 010109 99 138699 Expires 29 Oct 2013 Ready Figure 5 Controller image 8 way strip selection Ladder Needle Change Transport About Admin
22. es For total RNA samples Within a ScreenTape 18 2 Signal noise gt 3 in water and TE Agilent 2200 TapeStation User Manual Specifications 2 Specification P200 ScreenTape Assay Analytical Specification P200 ScreenTape and Reagents Sizing range Resolution Typical Sizing Accuracy Sizing Precision Quantitative Range precision Qualitative Range Sensitivity Physical Specification Sample volume needed Analysis Time Samples consumable Kit Size Kit Stability 10 200 kDa 15 96 10 CAII Lysozyme beta lactoglobulin 396 CV 100 1000 ng pL for IgG 15 CV 5 5000 ng L BSA Lysozyme 12 5 5000 ng yL IgG 5 ng pL Lysozyme 12 5 ng pL IgG 2 uL 16 samples 15 min 16 112 Samples 4 months 1 signal noise ratio gt 3 Agilent 2200 TapeStation User Manual determined using P200 Lader as sample 19 2 Specifications ScreenTape Specifications 20 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual e0 ee 3 e Installing the System Unpacking the System 22 Contents of the ScreenTape System 24 Installing the System 27 Software Installation 27 Agilent 2200 TapeStation Set Up 28 This chapter gives information about how to install the system RE Agilent Technologies 21 3 Installing the System Unpacking the System Unpacking the Agilent 2200 TapeStation Prerequisites Do not attempt to unpack the 2200 TapeStatio
23. fer 9 to 1 uL RNA sample or RNA Ladder 9 For best results use the reverse pipetting technique 5 Vortex using IKA vortexer and adaptor at 2000 rpm for 1 min 6 Spin down to position the sample at the bottom of the tube Agilent 2200 TapeStation User Manual 65 4 Using the 2200 TapeStation System RNA Sample Preparation 7 Sample denaturation a Heat samples to 72 C for 3 min b Place samples on ice for 2 min c Centrifuge to collect samples in the base of the tubes 1 uLRNA sample or Heat at 72 C ze RNA Ladder for 3 minutes W Place on ice for 2 minutes Vortex 1 min Spin down 66 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation High Sensitivity RNA ScreenTape Assay Prepare TapeStation Parts required p n Description 5067 5579 High Sensitivity RNA ScreenTape 1 Launch the Agilent 2200 TapeStation software 2 Load High Sensitivity RNA ScreenTape and loading tips into the 2200 TapeStation 3 Select RNA Protocol Eukaryotic RNA or Prokaryotic RNA Sample Preparation High Sensitivity RNA ScreenTape Assay Parts required p n Description 5067 5580 High Sensitivity RNA ScreenTape Sample Buffer 5067 5581 High Sensitivity RNA ScreenTape Ladder Allow reagents to equilibrate at room temperature for 30 min Vortex mix before use 1 2 3 Thaw total RNA samples on ice 4 Prepare Ladder solution by diluting to 20 uL with RNase free water 5 Add 1 uL Sample
24. g IKA vortexer and adaptor at 2000 rpm for 1 min 6 Spin down to position the sample at the bottom of the tube 3 pL D1000 Sample Buffer Vortex 1 min Spin down Agilent 2200 TapeStation User Manual 59 4 Using the 2200 TapeStation System Sample Preparation High Sensitivity D1000 ScreenTape Assay Parts required Parts required 60 2 uLDNA or Ladder High Sensitivity D1000 Prepare TapeStation p n Description 5067 5584 High Sensitivity D1000 ScreenTape 1 Launch the Agilent 2200 TapeStation software 2 Load High Sensitivity D1000 ScreenTape and loading tips into the 2200 TapeStation Sample Preparation High Sensitivity D1000 ScreenTape Assay p n Description 5067 5585 High Sensitivity D1000 Reagents High Sensitivity D1000 Ladder High Sensitivity D1000 Sample Buffer Allow reagents to equilibrate at room temperature for 30 min 2 Vortex mix before use 3 If running ladder add 2 uL High Sensitivity D1000 Sample Buffer 9 to 2 uL High Sensitivity D1000 Ladder C 4 Add 2 uL High Sensitivity D1000 Sample Buffer 9 to 2 uL DNA sample 5 Vortex using IKA vortexer and adaptor at 2000 rpm for 1 min 6 Spin down to position the sample at the bottom of the tube 2 uLD1000 Sample Buffer Vortex 1 min Spin down Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation Genomic DNA ScreenTape Assay Parts required Parts required Prepare TapeStation
25. ing the lid of the TapeStation after this time will abort the experimental run 44 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Running System WARNING Exposure to potentially dangerous mechanical parts gt Do not open the lid whilst the front LED is flashing Abort the TapeStation Run 1 If for any reason you wish to abort an experiment click the abort button on the pop up controller The instrument will ask a If you want to reset the instrument to begin another experiment this will return the controller software and TapeStation to the beginning of the next experiment b If you want to close down the controller this will close the controller software and keep the TapeStation temporarily locked in its current state Aborting the experiment will irretrievably discard any progress made and samples loaded Complete TapeStation Run When finished a pop up will ask for removal of the tip cartridge and tape 1 Remove tip cartridge and tape 2 Click OK Agilent 2200 TapeStation User Manual 45 4 Using the 2200 TapeStation System Empty Tip Buckets 1 Empty tip buckets Used loading tips must be removed from the tip buckets before the next experimental run The TapeStation will not start if tips are detected in the buckets Used ScreenTape sample strips and tips should be disposed of in accordance to local regulations How to Use the Agilent TapeStation Software For fu
26. must be taken due to viscosity of Sample Buffers When pipetting small volumes ensure that no sample remains within the tip When adding sample buffer to sample please ensure that they are mixed correctly by following assay instructions mproper mixing can lead to quantification errors Once mixed briefly centrifuge to collect the contents at the base of tubes Forbestresults ensure that all reagents are allowed to equilibrate to room temperature for 30 minutes prior to use When using 96 well plates the use of a 96 well plate vortex adaptor is advised to ensure correct sample mixing Improper mixing can lead to quantification errors As with samples in PCR strips briefly centrifuge after vortexing to collect the contents at the base of the tubes before placing into the TapeStation Agilent 2200 TapeStation User Manual 57 4 Using the 2200 TapeStation System Mixing recommendations TapeStation instruments are supplied with an optional IKA MS3 vortexer which includes a 96 well plate adaptor suitable for both 96 well PCR plates and 8 way strips This vortexer is recommended for use with the following applications D1000 ScreenTape Assay High Sensitivity D1000 ScreenTape Assay RNA ScreenTape Assay High Sensitivity RNA ScreenTape Assay Itis recommended that all current TapeStation users purchase this instrument direct from IKA for use with these assays Please quote part numbers 5067 5700 and 4674100 when ordering
27. n Analysis software However sizing may not be as accurate with a software ladder Ladders not run in the first available position or in No ladder mode can later be assigned as ladder using the 2200 TapeStation Analysis Software Agilent 2200 TapeStation User Manual 49 4 Using the 2200 TapeStation System Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation 50 Quantification Protocol Ensure that sample and sample buffer volumes are from the correct protocol Ensure that the reagents are used with the corresponding ScreenTape type Ensure correct pipetting technique When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Correct Mixing Sample and sample buffer must be vortex mixed according to protocol followed by centrifugation to remove any bubbles Insufficient mixing can cause discrepancies in quantification Mixing Protocols D1000 and High Sensitivity D1000 Vortex mix using IKA vortexer and adaptor at 2000 rpm for 1 minute Genomic DNA Manually vortex mix for 5 seconds on maximum speed Mixing Test 120 2 100 zz pig Soe 585 ome 40 e59 5 204 e Bud pum Recommended Protocol Pipette mixing only No mixing Figure 7 Effect of sample mixing on quantification results Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Peak Integration Ensure that the up
28. n instrument until you have read the accompanying Site and Safety Manual CAUTION Condensation within the instrument Condensation will damage the system electronics gt f your instrument was shipped in cold weather leave it in its box and allow it to warm slowly to room temperature to avoid condensation CAUTION Defective on arrival problems If there are signs of damage please do not attempt to install the instrument Inspection by Agilent is required to evaluate if the instrument is in good working condition gt Notify your local Agilent Representative and the Technical support channel gt An Agilent service representative will inspect the instrument at your site and initiate appropriate actions Personal injury The TapeStation is heavy gt Enlist the aid of a co worker to share the lifting load to avoid personal injury 22 1 Remove the TapeStation from the packaging and place on a clean dry flat surface 2 Allow the TapeStation to acclimatise to the ambient temperature of the operating environment Agilent 2200 TapeStation User Manual Installing the System 3 3 Remove the label covering the tape holder as shown in the image below Figure 4 Remove before use Delivery Checklist Ensure all parts and materials have been delivered with your system The delivery checklist is shown below Please report any missing or damaged parts to your local Agilent Technologies sales and service office
29. ng the most up to date Controller software Please check for the latest version You will now see the startup splash TapeStation Software TX Agilent Technologies 2 Insert the tube strip sample block into the TapeStation a Insert the plate sample block into the TapeStation E 36 Agilent 2200 TapeStation User Manual CAUTION Using the 2200 TapeStation System 4 3 Place loading tips into the loading tip holder as shown and insert into the TapeStation If any used tips are left in the tip buckets a pop up window will ask for the discarded tips to be removed The 2200 TapeStation will not run until all the tip buckets are empty A Loading tip transfer tool G2964 60000 is available Ensure that all 16 loading tips are inserted into the tip holder The laptop utilised for performing any previous use s of the ScreenTape must be utilised for all further re use Damage to the 2200 TapeStation and impact on performance Failure to use the correct consumable components can cause damage to the instrument gt Use the recommended loading tips gt Use the recommended foil plate seal 4 Remove ScreenTape from the foil packet Agilent 2200 TapeStation User Manual 37 4 38 Using the 2200 TapeStation System 5 Hold the tape with the ScreenTape label facing you and gently flick the top of the tape If there are any small bubbles present then this will move them to the top of the chamber
30. nt 5067 5576 RNA ScreenTape 7 ScreenTape 5067 5577 RNA ScreenTape Sample Buffer 1 vial Q 600 pL 5067 5578 RNA ScreenTape Ladder 1 vial o 10 uL Kit Components P200 ScreenTape Assay Part Number Name Color Amount 5067 5371 P200 ScreenTape 7 ScreenTape 5067 5372 P200 Reagents P200 5X Labeling Dye e 70 uL P200 Labeling Buffer e 350 uL P200 Reducing Sample Buffer Q 550 uL P200 pH Buffer clear 1000 uL P200 Non Reducing Sample Buffer 550 uL P200 Markers pre stained o 270 uL P200 Ladder e 40 uL 26 Agilent 2200 TapeStation User Manual Installing the System 3 Installing the System Software Installation The software for your Agilent 2200 TapeStation system is preinstalled on the system laptop For updates or if you have to change the laptop you may download the latest version of the software from the update server http www agilent com genomics tapestation For details on installation of the software refer to the readme txt file on the installation CD Agilent 2200 TapeStation Software TAPESTATION INSTRUMENT CONTROL AND DATA ANALYSIS Agilent 2200 TapeStation User Manual 27 3 Installing the System Agilent 2200 TapeStation Set Up Hardware required Software required Laptop Agilent 2200 TapeStation Software already installed Personal injury explosion or fire Do not operate the instrument in an atmosphere containing explosive gases or near flammable volatile liquids Only approved mains cord set
31. o the minimum required for the analysis CAUTION Damage to the 2200 TapeStation instrument gt Use only the recommended consumables and reagents with the 2200 TapeStation system When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to viscosity of Sample Buffers When pipetting small volumes ensure that no sample remains within the tip When adding sample buffer to sample please ensure that they are mixed correctly by following assay instructions mproper mixing can lead to quantification errors Once mixed briefly centrifuge to collect the contents at the base of tubes Forbestresults ensure that all reagents are allowed to equilibrate to room temperature for 30 minutes prior to use 48 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 For successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape Ladder Options In Ladder mode in the controller software a ladder should be loaded into the first available lane Alternatively the user can choose to run a software ladder This is done by choosing No ladder in the 2200 TapeStation Controller software ladder menu then running the instrument as normal A software ladder can then be inserted in the 2200 TapeStatio
32. p n Description 5067 5365 Genomic DNA ScreenTape 1 Launch the Agilent 2200 TapeStation software 2 Load Genomic DNA ScreenTape and loading tips into the 2200 TapeStation Sample Preparation Genomic DNA ScreenTape Assay p n Description 5067 5366 Genomic DNA Reagents 1 Equilibrate all reagents to room temperature for 30 min 2 Prepare Ladder a Aliquot a minimum of 3 uL Genomic DNA Ladder into the first tube well Use a fresh ladder for each run If using 96 well plates always run the ladder in first selected position No software ladder is available for the Genomic DNA assay Do not shake ladder vial This could result in degradation of the Genomic DNA ladder 3 Prepare Sample a Mix 1 uL genomic DNA sample 10 100 ng uL with 10 uL Genomic DNA Sample Buffer 9 b Spin down then vortex on maximum speed for 5 s c Spin down to position the sample at the bottom of the tube Agilent 2200 TapeStation User Manual 61 4 Using the 2200 TapeStation System DNA Sample Preparation 4 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section Tul 10 uL genomic Genomic DNA DNA Sample Buffer Genomic gt DNA Vortex on max am speed for 5 seconds Spin down 62 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 RNA Sample Preparation CAUTION Sample degradation gt Ensure all working areas reagents and plastic ware
33. per marker is properly integrated This is used for quantification Sample peaks should also be adjusted as required see Figure 8 on page 51 Sample A1 correct Sample A1 incorrect peak integration peak integration Concentration 42 2 ng uL Concentration 29 9 ng uL Be Pe Figure 8 Peak integration examples for correct and incorrect integration Quantitative Range For accurate quantification ensure that the sample is within the range of the chosen ScreenTape The quantitative range for D1000 ScreenTape is 0 1 50 ng uL The quantitative range for High Sensitivity D1000 ScreenTape is 10 1000 pg uL In extreme cases overloading the ScreenTape will result in a loss of the bottom marker Agilent 2200 TapeStation User Manual 51 4 Using the 2200 TapeStation System Other Issues Residual AMPure beads from SureSelect protocol can give signal which runs with the upper marker see figure below Any signal under the upper marker affects quantification Removal of the beads removes the signal under the upper marker This can be achieved using a magnetic plate as detailed in the Sure Select protocol If you see this signal please increase the duration of this step Enlarged image of upper m s marker with signal from AMPure beads ampio ptos Ful Figure 9 Upper marker with signal from AMPure beads Over amplification can also cause signal to run concurrently with the u
34. ple Preparation RNA ScreenTape Assay 65 Sample Preparation High Sensitivity RNA ScreenTape Assay 67 ah Agilent Technologies 31 4 Using the 2200 TapeStation System Protein Sample Preparation 69 Sample Preparation P200 ScreenTape Assay 70 This chapter explains the intended use of the 2200 TapeStation System 32 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Intended Use of the 2200 TapeStation System The 2200 TapeStation system Agilent 2200 TapeStation Software carries out electrophoretic separation of Nucleic Acids and proteins The system detects Fluorescently stained double stranded DNA including genomic DNA Fluorescently stained total RNA Eukaryotic and Prokaryotic Fluorescently labelled proteins Agilent 2200 TapeStation User Manual 33 4 Using the 2200 TapeStation System Performance Limitations of Use The 2200 TapeStation System Agilent 2200 TapeStation Software can analyse a maximum of 16 samples at any one time more samples can be run using a 96 well plate and multiple ScreenTape The user is responsible for establishing performance characteristics necessary for upstream and downstream applications Appropriate controls must be included in any upstream application requiring analysis on the 2200 TapeStation System Agilent 2200 TapeStation Software 34 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Additional Components Required by the User
35. pper marker 52 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sizing Analysis software mode Within the 2200 TapeStation analysis software sizing can be found in both electropherogram and region mode The sizing methods used in electropherogram and region mode will provide different sizing information Electropherogram View Region View Calculates data for a peak Calculates data for a smear or region The size reported is that of the highest point Size given is that of the centre of the in the peak region s mass Lower and upper marker Always ensure that the upper and lower markers have been identified correctly The markers are used as internal references to determine the molecular weight size of the sample Incorrect identification will lead to miscalculations in reported sizing values Agilent 2200 TapeStation User Manual 53 4 54 Using the 2200 TapeStation System Genomic DNA Equilibrate Reagents to Room Temperature Reagents must be equilibrated to room temperature for 30 minutes before use Failure to do so can affect sizing results Table 2 1500 1000 Sample Intensity FU Sample 17 k 50 ng uL Size Reagents at 4 C purple 24369 bp Genomic DNA Effect of temperature on sizing results MW bp Reagents Difference equilibrated to room temperature green 19867 bp 4502 bp Use f
36. quipment Directive 82 Technical Service 83 This chapter provides addition information RE Agilent Technologies 79 Appendix Limited Use Label License 80 These products may contain either SYBR Green or SYBR Gold nucleic acid stains which are provided under an agreement between Molecular Probes Inc a wholly owned subsidiary of Life Technologies Corporation and Agilent Technologies UK Limited and the manufacture use sale or import of this product is subject to one or more of U S Patents and corresponding international equivalents owned by Molecular Probes Inc The purchase of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer in conjunction with Agilent Technologies UK Limited s automated gel electrophoresis system where such research does not include testing analysis or screening services for any third party in return for compensation on a per test basis The buyer cannot sell or otherwise transfer a this product b its components or c materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes Commercial Purposes means any activity by a party for consideration and may include but is not limited to 1 use of the product or its components in manufacturing 2 us
37. r to use When using 96 well plates the use of a 96 well plate vortex adaptor is advised to ensure correct sample mixing Improper mixing can lead to quantification errors As with samples in PCR strips briefly centrifuge after vortexing to collect the contents at the base of the tubes before placing into the TapeStation For successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape For best sizing precision and accuracy run the appropriate ladder with the samples If 16 samples need to be analysed in parallel you may choose to insert a software ladder in the 2200 TapeStation analysis software However sizing may not be as accurate with a software ladder No Software ladder is available for the Genomic DNA application Agilent 2200 TapeStation User Manual 69 4 Using the 2200 TapeStation System Sample Preparation P200 ScreenTape Assay Parts required Parts required 70 Protein Sample Analysis p n Description 5067 5371 P200 ScreenTape 5067 5372 P200 Reagents 1 Launch the Agilent 2200 TapeStation software 2 Load P200 ScreenTape and loading tips into the 2200 TapeStation Sample Preparation P200 ScreenTape Assay p n Description 5067 5372 P200 Reagents 1 Prepare the P200 stain solution a Dilute P200 5X Labeling Dye 9 at a ratio of 1 5 with P200 Labeling Buffer 99 Th
38. resh ladder e Ladder must be prepared fresh for each run and run in the first available position Run profile decreases as ladder warms up and evaporates in the instrument e Will affect sizing results No software ladder will be available for genomic DNA Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 SENSU MU SYL lu ness ld eat ld A Ld e LS d Shaking Effect on Ladder If ladder is kept in the instrument over time the run profile of the lower marker changes e Manually shaking Genomic DNA Ladder vial by repeatedly inverting the tube results in degradation of the lambda DNA fragment This effect is not seen with vortex mixing AAA AT b A AA AWWW Tal ol al al alol alalla li Agilent 2200 TapeStation User Manual No shaking Shake for 40 seconds 55 4 Using the 2200 TapeStation System Molarity Molarity Molarity is determined from both size and quantity Errors in sizing and quantification will result in erroneous molarity calculations Always ensure that the good measurement practices for sizing and quantification have been followed to ensure accurate molarity values 56 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 DNA Sample Preparation When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care
39. rther information please refer to the software help This can be accessed by selecting the question mark button in the top right hand corner of the 2200 TapeStation Analysis Software 46 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Shutdown and Restarting Procedure Shutdown Procedure The controller software TapeStation instrument and laptop should be shut down when not in use preferably at the end of every working day Ensure that the TapeStation System is shut down in the following order 1 Exit the TapeStation Controller Software 2 Turn off the TapeStation instrument 3 Power down the laptop Restarting Procedure Ensure that the ScreenTape System is restarted in the following order 1 Power up the laptop 2 Turn on the TapeStation 3 Start the TapeStation Controller Software Agilent 2200 TapeStation User Manual 47 4 Using the 2200 TapeStation System How to prepare your samples KITE The handling of solvents samples and reagents can hold health and safety risks gt When using handling the ScreenTape and working with these substances observe appropriate safety procedures for example by wearing goggles safety gloves and protective clothing gt Always follow good laboratory practices and adhere to the guidelines established in your laboratory Refer to product material safety datasheets for further information gt The volume of substances should be reduced t
40. rument controller software is initialized and whilst the instrument is in operation LED The LED will illuminate once the instrument is on When the LED is flashing slowly the instrument is in use and the lid should not be opened rapid flashing indicates that the TapeStation requires some attention Sample Block There are 2 sample blocks provided that can either hold 0 2 mL sample tube strips or a 96 well plate Tip Holder The tip holder can accommodate up to 16 TapeStation loading tips at any one time ScreenTape The tape must be placed into the holder with the barcode towards the front of the instrument facing towards the right USB Socket The USB connector is inserted into the USB socket to link the laptop to the 2200 TapeStation Power cable socket The power cable must be connected to the 2200 TapeStation and the relevant mains electricity outlet 8 Agilent 2200 TapeStation User Manual Introduction to the 2200 TapeSt Lid ScreenTape Tip holder Sample block LED Figure 2 2200 TapeStation front view On off switch Power cable socket USB socket Figure 3 2200 TapeStation back view Agilent 2200 TapeStation User Manual ation System 1 1 Introduction to the 2200 TapeStation System Overview of the System 10 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual 9 RE ee e 00 2 Qeu9 Specifications e e e e Technical Specifications 12
41. s Agilent 2200 TapeStation User Manual Specifications 2 Specification RNA ScreenTape Assay Analytical Specification RNA ScreenTape and Reagents Quality Score Quantitative Range Quantitative Precision 96CV Quantitative Accuracy Sizing Range Sensitivity Analysis Type Maximum sample buffer strength Physical Specifications Analysis Time Samples per consumable Sample volume required uL Kit Stability Kit Size RIN 25 500 ng pL 5 20 100 6000 nt 5 ng pL Eukaryotic or Prokaryotic Total RNA QC 200 mM Tris 20 mM EDTA or 50 mM NaCl 16 samples lt 16 min 96 samples lt 100 min 16 1 4 months 112 samples For total RNA samples Within a ScreenTape Agilent 2200 TapeStation User Manual Signal noise gt 3 in water and TE 17 2 Specifications Specification High Sensitivity RNA ScreenTape Assay Analytical Specification RNA ScreenTape and Reagents Quality Score Quantitative Range Quantitative Precision 96CV Quantitative Accuracy Sizing Range Sensitivity Analysis Type Maximum sample buffer strength Physical Specifications Analysis Time Samples per consumable Sample volume required uL Kit Stability Kit Size RIN 500 10000 pg uL 10 30 100 6000 nt 100 pg pL Eukaryotic or Prokaryotic Total RNA QC 10 mM Tris 1 mM EDTA 16 samples 30 min 96 samples 180 min 16 2 4 months 112 sampl
42. scard this electrical electronic product in domestic household waste Product Category With reference to the equipment types in the WEEE Directive Annex I this product is classed as a Monitoring and Control Instrumentation product E Do not dispose off in domestic household waste To return unwanted products contact your local Agilent office or see www agilent com for more information 82 Agilent 2200 TapeStation User Manual Appendix 6 Technical Service For more information please contact Agilent Technologies UK Limited e www agilent com genomics contact Agilent 2200 TapeStation User Manual 83 Index Index A analysis mode select 36 C change needle 75 check final 44 condensation 22 D delivery checklist 23 E electronic waste 82 L limitations 34 load screentape 36 tips 36 needle change 75 notes add 41 P parts damaged 23 missing 23 84 protocol pipetting 50 sample and buffer 50 ScreenTape type 50 Q quantification 50 S samples describe 42 select samples 36 test 36 service technical 83 set up TapeStation 28 shutdown procedure 47 software startup 36 sound emission 81 specifications 11 Screenlape 14 T TapeStation abort 45 complete 45 setup 28 stat 44 unpacking 22 use 36 technical service 83 tip buckets empty 46 U unpacking 22 user change 36 use TapeStation 36 WwW waste electrical and electronic equipment
43. scriptions Sample Well Description D1000 ScreenTape ID 00 S029 010103 99 138100 Expires 29 Oct 2013 If the quoted current temperature is above the specified range please move the system out of direct sunlight and away from any windows Check that any air conditioning is functioning If the quoted current temperature is below the specified range please allow the instrument to equilibrate to the ambient temperature and avoid using in a cooled area Agilent 2200 TapeStation User Manual 13 2 Specifications ScreenTape Specifications Specification D1000 ScreenTape Assay Analytical Specifications D1000 ScreenTape and Reagents Sizing Range Typical Resolution Sensitivity Sizing Precision Sizing Accuracy Quantitative Precision Quantitative Accuracy Quantitative Range Physical Specifications Analysis Time Samples per consumable Sample volume required Kit stability Kit size 35 1000 bp 35 300 bp 15 300 1000 bp 10 0 1 ng pL 596 CV 10 0 1 1 ng uL 15 CV 1 50 ng pL 10 CV 20 0 1 50 ng uL 16 samples 20 min 96 samples 2100 min 16 1 uL 4 months 112 samples 1 signal to noise gt 3 single peak Sizing Accuracy for software ladder 20 3 Measured against 2100 Bioanalyzer 14 Agilent 2200 TapeStation User Manual Specifications 2 Specification High Sensitivity D1000 ScreenTape Assay Analytical
44. sult in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated condi tions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly performed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indi cated conditions are fully under stood and met Agilent 2200 TapeStation User Manual Contents Contents 1 Introduction to the 2200 TapeStation System 5 Overview of the System 6 2 Specifications 11 Technical Specifications 12 ScreenTape Specifications 14 3 Installing the System 21 Unpacking the System 22 Contents of the ScreenTape System 24 Installing the System 27 4 Using the 2200 TapeStation System 31 Intended Use of the 2200 TapeStation System 33 Performance Limitations of Use 34 Additional Components Required by the User 35 Operating Procedure 36 How to prepare your samples 48 Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation 50 DNA Sample Preparation 57 RNA Sample Preparation 63 Protein Sample Preparation 69 5 Maintenance 73 General Information 74 Changing the Needle 75 Agilent 2200 TapeStation User Manual Contents 6 Appendix 79 Limited Use Label License 80 Sound Emission 81 The Waste Electrical and Electronic Equipment Directive 82 Technical Ser
45. supplied with the instrument must be used with this instrument and if an extension lead is required the lead must be earthed gt lf the equipment is used in a manner not specified by the manufacturer the protection provided by the equipment may be impaired For general safety information please refer to the 2200 TapeStation System Site and Safety Manual Use of unsupplied cables or power adaptors Using cables or power adaptors not supplied by Agilent Technologies can lead to damage of the electronic components or personal injury gt Never use cables or power adaptors other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations 28 1 Connect the supplied USB cable between the port on the back of the instrument and your laptop 2 Power the instrument with the supplied power lead and adaptor 3 Turn the instrument on using the power switch located at the back of the TapeStation When powered and idle the instrument will have a blue LED visible on the front of the case Agilent 2200 TapeStation User Manual Installing the System 3 4 Windows may display a Found New Hardware wizard once the software has loaded In this instance always perform the following steps a Select No not this time to prevent connecting to Windows Update and searching for software b In the next window select Install the Software automatically c If a window appears
46. to purchase the correct needle cartridge Product Number TapeStation Configuration Pump Needle Cartridge Ordering Code ST007 TapeStation for ScreenPlex ST008 TapeStation for DNA Single G2960 60062 ST009 TapeStation for Nucleic acids ST017 TapeStation for ScreenPlex ST019 TapeStation for Nucleic acids Twin G2960 60063 STO10 TapeStation for Protein Combined TapeStation G2960A 2200 TapeStation System G2961A 2200 TapeStation Nucleic Acid System G2964AA 2200 TapeStation System Twin G2960 60063 G2965AA 2200 TapeStation Nucleic Acid System G2966AA 2200 TapeStation ScreenPlex System Agilent 2200 TapeStation User Manual 75 5 Maintenance Needle change intervals After 3840 7680 lanes in a Dual loading system pierces the controller software will inform the user that a needle change is pending The word Needle will appear in the bottom of the controller software inside a yellow box After 4160 pierces 8320 lanes in a Dual loading system a needle change is recommended The box around the word Needle will change from yellow to red After 4480 pierces 8960 lanes in a Dual loading system the needle has completed its lifetime and must be changed before the TapeStation will start leedle Change Transport Help TapeStationUser Agilent Technologies Sample Well Description Sample Descriptions Please insert a ScreenTape Ready Figure 10 Controller software indicating a Needle ch
47. user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or as a commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or dis closure of Software is subject to Agilent Technologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could re
48. vice 83 4 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual e e ee 1 e Introduction to the 2200 TapeStation e System Overview of the System 6 ScreenTape Architecture 7 Agilent 2200 TapeStation Components 8 This chapter gives an introduction to the system E Agilent Technologies 1 Introduction to the 2200 TapeStation System Overview of the System The Agilent 2200 TapeStation system is a revolutionary tape based platform for simpler faster and more reliable electrophoresis It is made up of three elements a consumable tape ScreenTape an instrument the 2200 TapeStation and an analysis software The system is very straightforward to use simply place the sample tubes and ScreenTape in to the 2200 TapeStation and let it load separate image analyse and present the results Place ScreenTape and some tips in the 2200 TapeStation Place your samples in the 2200 TapeStation and click Start on the controller software View your analysed results in around 1 min per sample This User Manual guides the operation of ScreenTape the 2200 TapeStation and software for the analysis of DNA RNA and protein samples The contents of the ScreenTape System are detailed below Information pertaining to the 2200 TapeStation can be found in 2200 TapeStation Technical Specification 2200 TapeStation Components Installing the 2200 TapeStation Agilent 2200 TapeStation User Manual
49. which have been run previously will appear grey For best sizing precision and accuracy the user should run the appropriate ladder with the samples If 16 samples are to be analysed in parallel the user may insert a software ladder for each application in the 2200 TapeStation analysis software However sizing may not be as accurate with a software ladder No software ladder is available for genomic DNA applications ScreenTape can be used up to 2 weeks after first use if it has been stored upright between 2 8 C Simply select the samples in the same manner as whole ScreenTape The first sample selected will automatically appear in the first available lane Partially used ScreenTape those that contain lanes run on previous occasions should be returned to the box and stored vertically between 2 8 C for a maximum of 2 weeks DNA Reagents Store from 2 8 C RNA Sample Buffer Store from 2 8 C RNA Ladder Store below 20 C P200 Reagents Store from 30 to 20 C Agilent 2200 TapeStation User Manual 39 4 Using the 2200 TapeStation System 10 The sample selection can be deleted by right clicking on the sample plate image A menu will appear with the following options Clear All Selections this will clear ladder well and all sample wells selected Clear Last Selection this will only clear the last samples to be highlighted Pressing Escape on the keyboard will also cancel the current s

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