Mondrian™ SP+ Cartridge
Contents
1. 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 M01344 v12. If you encounter any resistance to the cartridge moving into the deck inspect for foreign objects that may be 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 be slightly flexible in order to engage the cartridge However do not press too hard on the heater bars or Peltier unit as they may become misaligned Confirm that the cartridge is fully inserted into the deck When the cartridge is properly inserted the electrodes are no longer visible Figure 3 and the three guide arrows etched into the front of the deck should be flush with the edge of the cartridge Figure 4 Ill Operation 10 Mondrian SP Cartridge Figure 3 Inserting the Mondrian SP Cartridge A shows a cartridge improperly inserted into the workstation Note the visible electrodes B shows a cartridge correctly inserted A Figure 4 A close up view of the cartridge alignment marks etched into the deck of the Mondrian SP Workstation A no cartridge is inserted The alignment marks are clearly visible B the cartridge is correctly inserted The points of the guide arrows align with the edge of the cartridge A 5 Pull the locking lever forward as illustrated below to engage the control electronics and lock the cartridge into place Ill Opera
3. and then inserted into the deck of the workstation where a run is initiated through the touch screen 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 protocol performance the cartridge loading procedure must be strictly followed Consult this Mondrian SP Cartridge user guide and the appropriate NuGEN SP Library Systems user guide for the proper cartridge loading and setup procedure to achieve optimum results Figure 2 Top view of a Mondrian SP Cartridge Sample ports along the bottom edge are labeled S1 8 Other reagent port rows are labeled A E Note that there is no row B There are eight ports in rows A and C and seven in rows D and E The port labeled F is sized for a standard Luer lock connector through which the Filler Fluid is added to the cartridge The cartridge electrodes are visible as rows of small gold squares at the top of the cartridge E Reagents and master mixes F Filler Fluid port D Reagents and master mixes C Collection of final purified libraries A Adaptor ports Bubble trap ports nothing is added or removed via these ports Sample input ports 3 Il Kit Components Mondrian SP Cartridge The Mondrian SP Cartridge is shipped as part of a NUGEN SP Library System such as the Ovation SP Ultralow Library Systems Part Nos 8133 and 8134 or the Encore SP Complete Library Systems Part Nos 8
4. of 20 uL Depress the plunger on the pipette and insert the tip all the way to the bottom of the sample collection port perpendicular to the cartridge so as to make a seal between the cartridge and the pipet tip 3 Maintaining the seal formed between the pipette tip and the bottom of the cartridge release the plunger Immediately lift the pipette slightly off the bottom of the cartridge to release the seal rapidly 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 5 Dispense the collected sample into the sample vessel as directed in the user guide for your particular protocol If no sample droplet is observed dispense the contents of the pipette tip into the sample collection vessel and repeat the sample collection procedure for the port Note 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 6 Remove the cartridge from the workstation and dispose of in accordance with local regulations for laboratory waste IV Technical Support For help with any of our products please contact NUGEN Technical Support at 650 590 3674 direct or 888 654 6544 option 2 toll free U S only You may also send faxes to 888 296 6544 toll free or email techserv nuge
5. 151 and 8152 These kits contain all of the necessary reagents for complex sample preparation protocols on the Mondrian SP Workstation Each kit also contains protocol specific cartridge loading guides which can be overlaid on the cartridge to help direct the addition of reagents and samples Ill Operation The SP Filler Fluid has been degassed and pack 4 aged in a sealed single use vial Do not open the vial until ready for use Mondrian SP Cartridge A Powering up the Mondrian SP Workstation 1 Locate the instrument ON OFF switch at the back of the instrument and turn ON 2 Press the On button pictured below on the front of the instrument to power up the workstation B Filling the Cartridge SP Cartridges must be filled with the provided Filler Fluid prior to loading with reagents This process can be carried out on the bench top or the cartridge can be inserted into the Mondrian SP Workstation prior to adding filler fluid reagents and samples 1 Use scissors to cut open the Mondrian SP Cartridge pouch Remove the Mondrian SP Cartridge Note Do not attempt to tear open the pouch as this may damage the SP Cartridge 2 Lay the cartridge on a level bench top and place the appropriate SP Library Systems Cartridge Loading Guide on the cartridge 3 Remove the empty Filler Fluid tube and the SP Filler Fluid bottle from the NuGEN SP Library System kit and set both items on the bench top Il
6. Mondrian SP Cartridge FOR USE WITH PART NOS 8100 8133 8134 8151 AND 8152 Developed and Manufactured for NUGEN by Advanced Liquid Logic ge NuGEN imagine more from lesse Patents Licensing and Trademarks 2013 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 nugeninc 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 is covered by certain patents and trade secrets of Advanced Liquid Logic Inc licensed to NuGEN Inc Sale of the Mondrian SP Cartridge to NuGEN Inc customers comes with a limited research use only sublicense to use the Mondrian SP 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 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 transferable right without t
7. Q2 Can the Mondrian SP Cartridges be used with the Mondrian SP Workstation Yes The Mondrian SP Workstation is fully backward compatible and will run all NuGEN SP protocols Q3 Does NuGEN sell the Mondrian SP Cartridges as a stand alone No Mondrian SP Cartridges can be purchased only as part of a NuGEN SP Library System V Appendix ie NuGEN imagine more from less O4 Iuse the Library Prep Method 1 protocol on the SP Workstation and want to upgrade to the SP Workstation Can continue to use this protocol Yes The Mondrian SP Workstation supports the Library Prep Method 1 protocol on Mondrian SP Universal Cartridges Please visit www nugeninc com mondriansupport for more information and to download the correct protocol file for the Mondrian SP Workstation 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 uL custserv nugeninc com Fax 31 13 5780216 techserv nugeninc com europe nugeninc com 2013 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 nugeninc com NuGEN Ovation SPIA Ribo SPIA
8. duct eta rt EP E tudin 15 WMV Technical Support MM 16 LAE DD em G 17 A Cleaning the Workstation Contact Pins ssssssseeee 17 MM Ore ES 17 1 Introduction Mondrian SP Cartridge A Background The Mondrian SP Cartridge is used with the Mondrian SP System and offers a convenient 8 sample batch size simple reagent loading and easy sample recovery Reagents are contained in discrete droplets encased within filler fluid isolating the reactions from the lab environment and carryover contamination B Construction When prepared for use cartridges consist of an oil layer Filler Fluid sandwiched between a PCB substrate and a clear top plate The PCB substrate is patterned with insulated electrodes 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 sche matic of a cartridge Figure 1 Cartridge cutaway side view Top plate e d Droplet Filler fluid oil e a PCB substrate Hydrophobic coating Insulating polymer Electrode 2 Introduction Mondrian SP Cartridge C Cartridges Cartridges are unpacked filled with Filler Fluid reagents and samples
9. ges 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 Bl Contents Introduction csias tee m o ER ES RRIA NT E ORV EE E EEREE EE 1 A Background eR ES 1 B Constrictor anie r E EE sep EE RAEE 1 EEG slc gane E O E E E E R E NT 2 ll Kit Components cccssccsssssscssecssesssceeeeesesssceeeesseseseeeecessseseesecessseecescesssenseees 3 Ill POAC ONS noie eio aan ys Ii davssawsseacavssasacenedovanbadeneavosasctansiovaasaears 4 A Powering up the Mondrian SP Workstation sss 4 B EillinditheCattridge eee Ba P eret repete ES 4 C Cartridge Ifsertlohicsss dies teer det iot ates thee edis uo E to seus 8 D Cartridge Quality Control Check itte treten 11 E Loading Samples and Reagents ssssssssssssseeeen 14 E Qollectiomof Finished Pro
10. he 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 prod uct and methods accompanying this user guide 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 consequential or incidental dama
11. l Operation 4 Carefully grasp the tab on the red aluminium seal of the SP Filler Fluid bottle and pull up and back to break the seal as shown below e i d gf R B R Fluid por b S PERROS uat N m 5 Pull the tab until it breaks completely and remove the aluminium seal from the bottle Leave the grey rubber stopper in place N 6 Remove the top cover from the empty Filler Fluid tube leaving it capped at the bot tom Set the cap aside Important Do not discard the top cover 5 Mondrian SP Cartridge IIl Operation 7 Remove the rubber stopper from the bottle of SP Filler Fluid 8 Making sure the Filler Fluid tube is capped at the bottom slowly pour the SP Filler Fluid into the empty Filler Fluid tube Be careful not to pour too quickly as this will introduce bubbles into the fluid Air bubbles entering the cartridge may interfere with droplet movement 9 Once the entire aliquot of SP Filler Fluid is added recap the Filler Fluid tube 10 Invert the Filler Fluid tube and gently twist off the bottom cap of the Filler Fluid tube exposing the Luer lock tip as shown below 6 Mondrian SP Cartridge Ill Operation 7 Mondrian SP Cartridge 11 Insert the tip of the Filler Fluid tube into the port marked Filler Fluid on the SP Cartridge while gently twisting the tube in a clockwise direction to seat the tip of the Filler Fluid tube into the cartridge port Remove the top c
12. ninc com In Europe contact NUGEN at 31 0 135780215 Phone or 31 0 135780216 Fax or email europe nugeninc com In all other locations contact your NuGEN distributor for technical support 16 Mondrian SP Cartridge V Appendix 17 Mondrian SP Cartridge A Cleaning the Workstation Contact Pins The pins on the workstation cartridge interface may become dirty and cause perfor mance issues If this happens you will need to clean the interface pins Materials e Lint free wipes such as Kimwipes or Super PolX 1200 Wipers Do not use cotton or any material that may leave particles behind e Isopropyl alcohol e Canned air Procedure 1 Turn the workstation off and unplug from the power source 2 Soakthe wipe in isopropyl alcohol 3 Firmly rub all pins with the wipe 4 Wait 2 minutes 5 Blow the area dry with canned air 6 Plug the workstation back in and turn it on Note We recommend cleaning the pins once a week after any prolonged storage of the workstation after a Filler Fluid spill or if the Mondrian SP Cartridge OC proto col returns a Mondrian SP Cartridge Failed or a Mondrian SP Cartridge Status Undetermined message B FAQs O1 Can the Mondrian SP Cartridges be used with the Mondrian SP Workstation No The Mondrian SP Cartridges can be run only on the Mondrian SP Workstation Running an SP Cartridge on an SP Workstation will result in a failed experiment and loss of samples
13. over from the Filler Fluid tube allowing the entire volume of Filler Fluid to flow into the cartridge This process will take approximately 3 5 minutes During the cartridge filling process you will begin to see the filler fluid rising in the sample and reagent ports You should see filler fluid in all sample and reagent ports by the end of the filling process Ill Operation 8 Mondrian SP Cartridge Full Not yet completed filling 12 Once the entire volume of Filler Fluid has been delivered carefully remove the 13 Filler Fluid tube from the SP Cartridge Inspect the cartridge to make certain that Filler Fluid has risen into each of the sample and reagent ports Occasionally some of the ports may fail to properly fill due to surface tension If this is the case the cartridge may be tilted slightly to encourage the Filler Fluid to enter the ports Alternatively a pipet tip may be used to break the surface tension in affected ports Cartridge Insertion Raise the lid of the Mondrian SP Workstation to reveal the cartridge deck Ill Operation 9 Mondrian SP Cartridge With the cartridge lever in the up position as shown above hold the cartridge level by the plastic top cover with ports facing up and the gold colored electrodes point ing 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
14. ple Input OOOQOQ000 P01210 Follow the recommendations below to load reagents into their appropriate cartridge ports Use a 10 or 20 uL pipette to add all reagent master mixes whose volume is less than 10 uL as well as the adaptors e Use a 100 or 200 uL pipette for adding any reagent of a volume greater than 20 uL Ill Operation 15 Mondrian SP Cartridge Pipet samples and reagents carefully to avoid overflow of the Filler Fluid from the port e When adding samples and reagents insert the pipette tip all the way to the bottom of the cartridge If the tip contacts the bottom withdraw the pipette tip slightly to allow space for dispensing Slowly depress the plunger to dispense the reagent but do not depress the plunger completely blow out as this could introduce bubbles into the cartridge Refer to the Mondrian SP Workstation User Manual and the appropriate NuGEN SP Library System user guide for guidance on protocol selection and starting the sample preparation run F Collection of Finished Product Note A short instructional video on Mondrian Sample Collection is available on the Mondrian Support page at www nugeninc com mondriansupport 1 Once the protocol run has completed raise the lid and prepare all necessary mate rials for sample collection from the cartridge as directed in the appropriate NuGEN SP Library System user guide 2 Use a 100 or 200 pL pipette and set it to a volume
15. re inconclusive Press OK on the Run Complete screen to return to the main menu Remove the cartridge from the deck and set it aside do not discard Clean the contact pins on the workstation following the protocol in Appendix C Cleaning the Workstation Contact Pins Repeat the Cartridge OC protocol from step 4 above one more time Do not add any additional Filler Fluid or Elution Buffer to the cartridge If the message after the second Mondrian SP Cartridge OC protocol is e Mondrian SP cartridge passed proceed as described for passing cartridges above Mondrian SP Cartridge failed or Mondrian SP cartridge status is undetermined do not use the cartridge Contact NuGEN Technical Support Ill Operation 14 Mondrian SP Cartridge E Loading Samples and Reagents Protocols for Mondrian SP products can be found in the product specific user guides which can be downloaded from www nugeninc com mondriansupport Many SP sample preparation protocols may appear to be similar so make certain you are work ing from the appropriate user guide Use the product specific SP cartridge loading guides to aid in proper placement of reagents and samples Figure 5 Figure 5 An example cartridge loading guide E2 E3 E4 E5 0000 OO D6 D7 5000668 Sample Colson 1QOOOOOO 0000060 Adaptors Ovation SP Ultralow DR Multiplex Systems Single use only ge NUGEN rc
16. rian SP Cartridge 9 If the cartridge is not already inserted into the Mondrian SP Workstation carefully transport the cartridge to the Mondrian SP Workstation and insert the cartridge into the deck Pull the cartridge lever of the Mondrian SP Workstation forward to the locked position and close the lid of the workstation Select Run on the touch screen menu choose the Mondrian SP Cartridge QC 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 in the Run Information screen We highly recom mend recording the serial number of the cartridge in the Run Information screen The cartridge serial number can be found on the front of each cartridge for easy reference Select Next and then select Start Run The Mondrian SP Cartridge QC protocol will take about 15 minutes to complete During this test Elution Buffer droplets will be dispensed from the E5 port and trans ported around the cartridge prior to being returned to the port The purpose of this test is to confirm the basic performance of the cartridge Ill Operation 13 Mondrian SP Cartridge At the end of the protocol the instrument will display the Run Complete screen and one of the following messages SP Cartridge failed Remove cartridge from instrument deck and set aside prior to contac
17. t ing NuGEN Technical Support detected with droplet trans port within the cartridge MESSAGE MEANING NEXT STEP Mondrian No errors were Press OK on the Run Complete screen to return SP Cartridge detected Droplet to the main menu Proceed to section D Loading passed transport was Samples and Reagents in this user guide or Continue normal Cartridge follow the instructions in the appropriate NuGEN to intended is ready to runa SP Reagent System protocol protocol Mondrian A problem was Press OK on the Run Complete screen to return to the main menu Remove the cartridge from the deck and set it aside do not discard Clean the contact pins on the workstation following the protocol in Appendix C Cleaning the Workstation Contact Pins Repeat the Cartridge QC protocol from step 4 above one more time Do not add any additional Filler Fluid or Elution Buffer to the cartridge If the message after the second Mondrian SP Cartridge OC protocol is e Mondrian SP cartridge passed proceed as described for passing cartridges above e Mondrian SP Cartridge failed or Mondrian SP cartridge status is undetermined do not use the cartridge Contact NuGEN Technical Support Mondrian SP Cartridge status is undeter mined Please consult the Mondrian SP Cartridge user guide or appro priate NuGEN SP Library Systems user guide for further instructions The results a
18. tion 11 Mondrian SP Cartridge Note Once a run has started do not move the locking lever as this will result in an unrecoverable interruption of the running protocol The Mondrian SP Workstation will not run with the lever in the up position D Cartridge Quality Control Check The Mondrian SP Cartridge OC protocol allows confirmation of the basic functionality of the Mondrian SP Cartridge prior to use We highly recommend running this proto col with each cartridge prior to adding samples and reagents The Cartridge OC loading process can be carried out on the bench top or the cartridge can be inserted into the Mondrian SP Workstation prior to adding filler fluid and elu tion buffer T Place the Cartridge Loading Guide provided with each Mondrian SP Cartridge on the cartridge Note It is possible to place the loading guide on the cartridge in only one orientation Remove the Elution Buffer reagent tube from the room temperature storage com ponents of a NuGEN SP Library System kit Load 50 uL of Elution Buffer into port E5 of the Mondrian SP Cartridge Insert the pipette tip into the port all the way to the bottom of the cartridge If the tip contacts the bottom withdraw the pipette tip slightly to allow space for dispens ing Slowly depress the plunger to dispense the reagent but do not depress the plunger completely blow out as this could introduce bubbles into the cartridge Ill Operation 12 Mond
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