Mondrian™ SP Universal Cartridge
Contents
1. NuGEN Technologies Inc Headquarters USA Europe 201 Industrial Road Suite 310 P O Box 109 For our international distributors contact San Carlos CA 94070 USA 9350 AC Leek information visit our website Toll Free Tel 888 654 6544 The Netherlands Toll Free Fax 888 296 6544 Tel 31 13 5780215 uico LE custserv nugen com Fax 31 13 5780216 techserv nugen com europe nugen com 2012 2015 NuGEN Technologies Inc All rights reserved The Encore Ovation and Applause families of products and methods of their use are covered by several issued U S and International patents and pending applications www nugen com NuGEN Ovation SPIA Ribo SPIA Applause Encore Prelude Mondrian and Imagine More From Less are trademarks or registered trademarks of NUGEN Technologies Inc Other marks appearing in these materials are marks of their respective owners For research use only M01265 v2 12. e nba toe ina ooo ER o eye rator Fo ra eas 14 MP DIPL E 15 A Troubleshooting uide ce teet e e rte t bebe E e dit ts 15 1 Introduction Mondrian SP Universal Cartridge A Background The Mondrian SP Universal Cartridge kit is provided for use with protocols that use sample preparation reagents from third party vendors and includes a single use disposable cartridge designed to facilitate hands free digital microfluidics genomic sample preparation on the Mondrian SP Workstation The SP Cartridge is used in all Mondrian SP protocols and offers a convenient 8 sample batch size simple reagent loading and easy recovery of sample Reagents are contained in discrete droplets encased within filler fluid isolating the reactions from the lab environment and carry over contamination B Construction Cartridges prepared for use consist of an oil layer Filler Fluid sandwiched between two substrates onto which insulated electrodes are patterned By changing the relative voltages of the patterned electrodes aqueous droplets are manipulated to perform complex assays Droplets are dispensed from loading ports transported to various locations on the cartridge mixed incubated and collected using only software control Figure 1 shows a cutaway side view schematic of a cartridge Figure 2 shows a top view of the cartridge Figure 1 Cartridge cutaway side view Top plate S lt Droplet Filler fluid oil e a Insulating polymer PC
3. the Mondrian SP Workstation Detailed instructions for loading reagents and samples are application specific Many sample preparation protocols may appear to be similar Verify that you are referring to the User Guide specific to the sample preparation protocol you intend to run and that you have obtained all of the necessary additional reagents and materials prior to loading the cartridge Contact NuGEN Technical Services at techserv nugen com for more information Ensure that samples and reagents are correctly loaded into the cartridge before start ing the run Use the application specific loading guides to aid you in proper placement of reagents and samples An example of the Library Preparation Method 1 loading guide is shown in Figure 4 Figure 4 An example cartridge loading guide Guide pictured is for Library Prepa ration Method 1 0005508 OOUOOOD COCCUSS OG Or O Q OO OQ OQ ar OOO OC Library Preparation Method 1 Single use only J gacen C9996009 When adding sample or reagent place the tip of the pipette below the surface of the filler fluid and slowly depress the plunger When adding reagents to the smaller ports labeled A B or C on the cartridge refer to Figure 2 above guide the pipette tip gently all the way to the bottom of the well Slowly depress the plunger to dispense the reagent completely and carefully pull the pipette tip back out Adding samples or reagents too quickly can cause the loading po
4. B substrate Electrode Hydrophobic coating 2 Introduction Mondrian SP Universal Cartridge C Cartridges Cartridges are unpacked inserted into the workstation loaded with filler fluid samples and reagents and a run is initiated through the touch screen user interface Cartridges are one time use only Cartridge re use is not supported even if a run is canceled prior to completion To ensure proper assay performance cartridge loading procedure must be strictly followed Consult this Mondrian SP Universal Cartridge User Guide and appropriate Application Note or Application User Guide for proper cartridge loading and setup procedure to achieve optimum results Figure 2 shows a diagram of a Mondrian SP Cartridge Figure 2 Top view of the cartridge Sample ports along the bottom edge are labeled S1 8 Other reagent port rows are labeled A E Note that there are 8 ports in rows A C and 7 in rows D and E There is a port labeled OIL sized for a standard Luer lock connector where the Filler Fluid is added to the cartridge The cartridge electrodes are visible at the top of the cartridge as rows of small gold squares ll Kit Components 3 Mondrian SP Universal Cartridge The Mondrian SP Universal Cartridge kit is shipped with ancillary consumables for use with third party reagents as detailed in the appropriate Application Note contact NuGEN Technical Services at techserv nugen com to perform complex sample prep
5. Mondrian SP Universal Cartridge PART NO 8010 Developed and Manufactured for NUGEN by Advanced Liquid Logic ge NuGEN imagine more from lesse Patents Licensing and Trademarks 2012 2015 NuGEN Technologies Inc All rights reserved The Encore Ovation and Applause families of products and methods of their use are covered by several issued U S and International patents and pending applications www nugen com NuGEN Ovation SPIA Ribo SPIA Applause Encore Prelude Mondrian and Imagine More From Less are trademarks or registered trademarks of NUGEN Technologies Inc Other marks appearing in these materials are marks of their respective owners The Mondrian SP Universal Cartridge is covered by certain patents and trade secrets of Advanced Liquid Logic Inc licensed to NuGEN Inc Sale of the Mondrian SP Universal Cartridge to NuGEN Inc customers comes with a limited research use only sublicense to use the Mondrian SP Universal Cartridge for preparing a nucleic acid sample for analysis on a separate system by sequencing microarray or real time PCR The sublicense specifically excludes the right to use the Mondrian SP Universal Cartridge for biothreat detection forensic testing production animal health testing and food safety testing except for genomics research in these fields and for human and veterinary diagnostics The purchase of this product conveys to the buyer the limited non exclusive non transferab
6. a ration assays on the Mondrian SP Workstation Protocol specific loading guides that can be overlayed on the cartridge are also provided Table 1 details the kit contents All components should be stored at room temperature Table 1 Additional Reagents and Components Supplied with Mondrian SP Universal Cartridge MONDRIAN SP UNIVERSAL CARTRIDGE KIT 8010 08 PART NUMBER 8010 32 PART NUMBER 1 cartridge of 4 cartridges of PRERSIER 8 samples cartridge 8 samples cartridge Reagent Additive 501555 501586 Sample Concentration Solution 501556 501587 Bead Binding Solution 501557 501588 Bead Wash Solution 501558 501589 Elution Buffer 501559 501590 Filler Fluid 501561 501561 SP Cartridge empty P01190 P01190 Cartridge Loading Guide s P01189 P01189 Ill Operation 4 Mondrian SP Universal Cartridge A Filling the Cartridge SP Cartridges must be filled with the provided Filler Fluid prior to loading with reagents Cut open the foil packet containing the cartridge remove the cartridge from the packet and place on lab bench Alternatively the cartridge can be inserted into the deck of the Mondrian SP Workstation and locked into place with the cartridge lever prior to filling and loading See Section III C for instructions on inserting the cartridge into the Mondrian SP Workstation Remove the tube of Filler Fluid from the kit box and gently twist off the bottom cover expos
7. bars or Peltier unit as this may cause them to become misaligned Ill Operation Figure 6 Inserting the cartridge 3 Confirm that the cartridge fits snugly into the rear of the deck and that the elec trodes are not visible from above Figure 6 This will ensure that the electrodes will make good contact with the control electronics at the rear of the deck When the cartridge is properly inserted three guide arrows etched into the metal surface at the front edge of the deck should be visible with their tips flush with the edge of the cartridge as illustrated in Figure 8 Figure 7 Determining if the SP Cartridge is properly loaded in the deck If the electrodes are still visible after the cartridge has been loaded A the cartridge loading has not been completed and the cartridge should be pushed further back until no electrodes are visible from above B A 00090 0000094 Sample Collection a 2 2 2 e B 10 Mondrian SP Universal Cartridge Ill Operation Figure 8 Gently insert the cartridge into the deck until the front edge of the cartridge is aligned with the tips of the machined arrows Do not stop inserting the cartridge when the front edge is flush with the edge of the deck A but continue to insert the cartridge until the edge aligns with the tips of the arrows B A Library Preparation Method 1 Single use only XE NUGEN magne more tom les gt Input esce Figure 9 Cartr
8. ed The purpose of this test is to confirm the successful transport of droplets across all lanes of the cartridge At the end of the protocol the instrument will display the Run Complete screen and one of the following messages Ill Operation MESSAGE MEANING NEXT STEP Mondrian No errors Press OK on the Run Complete screen to return SP cartridge detected to the main menu Proceed to section C Loading passed Droplet transport Samples and Reagents below in the Mondrian Continue was normal SP Universal Cartridge user guide or follow the to intended instructions for 3rd party reagents in the appropri protocol ate applications note The user may remove the cartridge from the Workstation to load samples and remaining reagents on the bench top taking care that the cartridge remains level at all times Note It is not necessary to add additional Elution Buffer or Filler Fluid to the cartridge prior to use Mondrian A problem was Press OK on the Run Complete screen to return SP cartridge detected with to the main menu Remove the cartridge from the failed Remove cartridge from instrument deck and set aside prior to contact droplet trans port within the cartridge deck and set it aside do not discard and contact NuGEN Technical Support for further instructions Users who proceed with SP protocols and or who load additional SP reagents and samples onto failed cartridges are doing so at their own risk and wi
9. f the finished products Dispose of the cartridge as appropriate in laboratory waste IV Technical Support For Technical Support please contact NuGEN at U S only 888 654 6544 Toll Free Phone or 888 296 6544 Toll Free Fax or email techserv nugen com In Europe contact NuGEN at 31 0 135780215 Phone or 31 0 135780216 Fax or email at europe nugen com In all other locations contact your NuGEN distributors Technical Support team 14 Mondrian SP Universal Cartridge V Appendix i NuGEN imagine more from less A Troubleshooting In the event of a performance issue with the Mondrian SP System the most likely cause is due to incorrect loading of the cartridge or the insertion of the cartridge into the deck of the Workstation Ensure that you follow directions carefully and ensure that all reagents and samples have been loaded into the correct locations Failed reac tions may also result from electrodes that have become fouled with Filler Fluid See the Mondrian SP Workstation User Manual document M01264 for instructions on cleaning of the electrodes For other issues please contact NuGEN Technical Support at techservOnugen com B Update History This document the Mondrian SP Universal Cartridge User Guide M01265v2 1 is an update to address the following topics DESCRIPTION VERSION SECTION PAGE S Update patent and copyright statement warranty statement and URLs Throughout Throughout
10. he pipette and insert the tip all the way to the bottom of the extraction port perpendicular to the cartridge Make sure that you make contact with the bottom of the cartridge 3 Maintain contact of the pipette tip with the bottom of the cartridge and release the plunger Lift the pipette off the bottom of the cartridge to release the vacuum that has been created drawing filler fluid and the sample droplet into the pipette tip 4 Examine the pipette tip to ensure that the appropriately sized droplet is sus pended in the Filler Fluid in the pipette tip Ill Operation 13 Mondrian SP Universal Cartridge Dispense the collected sample into the sample vessel as directed in the docu mentation for your particular protocol If you do not see the sample droplet in the pipette tip dispense the contents of the pipette tip into the sample collection ves sel and repeat the sample collection procedure for the port Sometimes it is neces sary to repeat the sample collection Note that the droplet occasionally breaks into pieces and becomes more difficult to observe The excess filler fluid collected during the procedure will not interfere with downstream molecular processes After all the finished products have been collected from the cartridge push the locking lever away from you to disengage the cartridge and remove the cartridge from the instrument deck by pulling it forward if the cartridge was not already removed prior to the collection o
11. ial or incidental damages arising from the use results of use or inability to use its products NuGEN reserves the right to change alter or modify any product to enhance its performance and design NuGEN s products are developed designed and sold FOR RESEARCH USE ONLY This product is not to be used for diagnostic or therapeutic purposes nor is it to be administered to humans or animals Except as expressly set forth herein no right to modify reverse engineer distribute offer to sell or sell NuGEN s product is conveyed or implied by buyer s purchase of this NuGEN prod uct The buyer agrees to use NuGEN products accompanying the product insert in accordance with the intended use and the written instructions provided Table of Contents Contents Introd ctiomnaucne tette m ER RS IRRA ONT EE E EESE 1 A BaEkGgr OU seed eisustenee t Da e a E AE E eases EEEURE E EEEN 1 B Constrictor anii r d rk Dres Rp ED REEE 1 EEG slc yare E ees die oatarcusesacassavess E E E R E N 2 ll Kit Components cccscccscssssssscessesssceseeesessseeeeesseseseesecessscsesecesssensceecessseneeees 3 ESI CUDDLEE M 4 As Eilling the Cartridge ete d a tepore ERE RENE EREE ONERE IBS 4 B Cartridge Quality Control Ghieck note er HE testet 4 C Loading Samples and Reagents sssssssssssssseeeneeeeeens 8 De e inufelei ibtii p 9 E Collectionvot Finished Product ate rtt an 12 WV Technical Support iere rer tri ena
12. idge inserted into deck 4 Pullthe locking lever forward and down as illustrated in Figure 10 to engage the control electronics and lock the cartridge into place 11 Mondrian SP Universal Cartridge Ill Operation 12 Mondrian SP Universal Cartridge Figure 10 Locking the cartridge deck Note Once a run has started do not raise the lever as this will result in an unre coverable interruption of the running protocol and an error flag The Mondrian SP Workstation will not run with the lever in the up position Note Filler Fluid addition and loading of the cartridge may take place either with an empty cartridge inserted into the Mondrian SP Workstation and locked into place or on the bench top with subsequent insertion of the cartridge into the Mondrian SP Workstation Once a fully loaded cartridge has been inserted into the Mondrian SP Workstation close the lid and proceed with running the desired protocol See the Mondrian SP Workstation User Manual and the appropriate Application Note or SP Reagent System User Guide for instructions on protocol selection and starting the sample preparation run E Collection of Finished Product 1 Once the protocol run has completed raise the lid and prepare all necessary materials for sample collection from the cartridge as directed in the appropriate Application Note or SP Reagent User Guide 2 Select a 100 or 200 uL pipette and set it to a volume of 20 uL Depress the plunger on t
13. ing the Luer lock tip Carefully insert the tip onto the port marked OIL on the cartridge while twisting the tube in a clockwise direction as you do this to firmly seat the cartridge on the port Once the tube is firmly seated such that it remains upright in place carefully snap off the top cover from the tube taking care that the Filler Fluid tube remains upright and connected to the cartridge Filler Fluid will begin to flow into the cartridge The Filler Fluid will come partway up the sample and reagent ports but will not overflow onto the cartridge as long as the cartridge remains level Carefully remove the Filler Fluid tube from the cartridge while holding the cartridge in place on the bench top and discard the empty Filler Fluid tube If some of the ports do not appear to have filler fluid partway up the port gently tap or very gently shake the car tridge on the bench top keeping the cartridge level and avoid spilling Filler Fluid out of the cartridge B Cartridge Quality Control Check The Mondrian SP Cartridge QC protocol is a QC check that we recommend be per formed prior to adding samples and reagents to the Mondrian SP Cartridge This QC check confirms the functionality of the Mondrian SP Cartridges prior to use The Mondrian SP Cartridge OC protocol requires inserting the Filler Fluid containing cartridge into the deck of the Mondrian SP Workstation Some customers may find it easier to place the fluid containing cartridge o
14. le right without the right to modify reverse engineer resell repackage or further sublicense under these patent applications and any patents issuing from these patent applications to use this product and methods accompanying this package insert for research and development purposes solely in accordance with the intended use described and the written instructions provided in this user guide No license to make or sell products by use of this product is granted to the buyer whether expressly by implication by estoppels or otherwise In particular the purchase of this product does not include or carry any right or license to use develop or otherwise exploit this product commercially and no rights are conveyed to the buyer to use the product or components of the product for purposes including commercial services or clinical diagnostics Warranty NuGEN warrants that this product meets the performance standards described in the Company s product and technical literature for a period of six months from the date of purchase provided that the product is handled and stored according to published instructions and that the product is not altered or misused If the product fails to meet these performance standards NuGEN will replace the product free of charge or issue a credit for the purchase price NUGEN s liability under this warranty shall not exceed the purchase price of the product NUGEN shall assume no liability for direct indirect consequent
15. ll not ing NuGEN be compensated for loss of reagents samples or Technical cartridges by NuGEN Technologies Inc Support Mondrian The results are Repeat the Mondrian SP Cartridge OC protocol one SP cartridge inconclusive more time status is and must be undetermined Please consult Mondrian SP Universal Cartridge User Guide or appropriate NuGEN SP Library Systems User Guide for further instructions repeated one more time prior to making a determination on the quality of the cartridge Do not add any additional Filler Fluid or Elution Buffer to the cartridge Press OK on the Run Complete screen to return to the main menu Proceed to re run the Mondrian SP Cartridge OC protocol starting from Step 10 in the protocol above If the message after the second Mondrian SP Cartridge OC protocol is Mondrian SP cartridge passed then proceed as outlined in Next Step for passing cartridges above Mondrian SP Cartridge failed or Mondrian SP cartridge status is undetermined do not use the cartridge and contact NuGEN Technical Support Mondrian SP Universal Cartridge Ill Operation 8 Mondrian SP Universal Cartridge C Loading Samples and Reagents Sample and Reagent addition can be performed either while the cartridge is resting on the bench top or in the deck of the Mondrian SP Workstation See Section III C for instructions on inserting the cartridge into
16. n the workstation and then pipet the Elution Buffer into the cartridge Alternatively it may be easier to pipet the Elution Buffer into the fluid containing cartridge while the cartridge rests on the bench top and then insert the reagent and fluid filled cartridge into the deck of the Workstation moving carefully to avoid spilling the fluid or disturbing the reagent 1 Locate the Cartridge Loading Guide that is provided with each Mondrian SP Cartridge and place the loading guide on the cartridge Note it is only possible to place the loading guide on the cartridge in a single orientation Ill Operation 5 Mondrian SP Universal Cartridge Optional Carefully move the Filler Fluid containing cartridge and its Cartridge Loading Guide to the deck of the Mondrian SP Workstation and insert the car tridge into the deck Remove the Elution Buffer reagent tube shipped and stored at room tempeera ture from the Mondrian SP Universal Cartridge kit e Part 501559 in the Mondrian SP Universal Cartridge 1 pack part 8010 08 or Part S01590 in the Mondrian SP Universal Cartridges 4 pack part 8010 32 Set a 50 or 100 uL single channel pipette to 50 pL and remove 50 uL of Elution Buffer from the tube Load 50 uL of Elution Buffer into port E5 of the Mondrian SP Cartridge When adding the reagent to the cartridge place the tip of the pipette below the surface of the filler fluid and slowly depress the plunger Dispense the reagen
17. rt or adjacent ports to overflow and create the potential for sample cross contamination Multichannel pipettes can be used and are recommended particularly for adding samples and reagents such as library adaptors to the sample ports and ports in rows A C P01189 v1 N 9 Ill Operation Mondrian SP Universal Cartridge D Cartridge Insertion 1 Raise the lid of the Mondrian SP Workstation to reveal the cartridge deck Figure 5 Figure 5 Top view of the cartridge deck of the Mondrian SP Workstation E 2 With the cartridge lever in the up position lifted and moved toward the rear of the deck hold the cartridge level with the plastic top cover with ports facing up and the gold colored electrodes pointing towards the Workstation Carefully insert the cartridge into the cartridge deck flush with the surface of the deck allowing the side rails to guide it into place see Figure 6 If you encounter any resistance to the cartridge moving into the deck carefully inspect for any foreign objects that are impeding the cartridge and remove them If the cartridge appears to be catching on the edge of the heater bars or Peltier cooling plate you may gently press down on them to allow the cartridge to move forward The heater bars and cooling unit are designed to move up and down slightly in order to firmly engage the cartridge when the cartridge lever is in the down and locked position However do not press too hard on the heater
18. t completely and carefully pull the pipette tip back out Note Do NOT add any samples or other reagents to the cartridge at this time Ensure that only Elution Buffer has been loaded If the cartridge is not already on the Mondrian SP Workstation deck carefully transport the cartridge to the Mondrian SP Workstation and insert the cartridge into the deck If not already ON locate the instrument ON OFF switch at the back of the instrument and turn ON Press the On button Figure 3 on the front of the instrument to turn the instrument on Figure 3 The Mondrian SP Workstation On button located on the front of the instrument 10 11 12 13 Pull the cartridge lever of the Mondrian SP Workstation forward to the locked posi tion and close the lid of the workstation Select Run on the touch screen menu choose the Mondrian SP Cartridge OC protocol from the list of protocols and then select Next to proceed to the Protocol Information screen Select Next to proceed to the Run Information screen Optional Enter run details as required in the Run Information screen Select Next to proceed to the Run Confirmation screen and select Start Run 6 Ill Operation Mondrian SP Universal Cartridge The Mondrian SP Cartridge OC protocol will take about nine minutes to complete During this test Elution Buffer droplets will be dispensed from the E5 reservoir and transported around the cartridge prior to being discard
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