EpiSonic - LAB MARK
Contents
1. m Do not discard the original packaging You may need the packaging materials in the future for servicing your device m The EpiSonic device contains sensitive electronics Always be gentle when removing parts from the packaging box or handling parts in general m To prevent any unnecessary failure in the fuse make sure to always turn off the power supply before connecting or disconnecting the power cord m Confirm that the power switch on the rear panel of the sonication generator is switched to the OFF position m Use only the power cord supplied by Epigentek You can also use an appropriate adapter for your specific country in the event the plug does not match your wall outlet The EpiSonic device uses a universal power supply that recognizes any AC mains voltage throughout the world I PRE ASSEMBLY Converter amp Horn Assembly Start by placing the Sample Pro cessing Horn upside down Care fully align the Converter to connect it to the Sample Processing Horn by the threaded stud hand tighten Next use the strap wrench in one hand to grip the Sample Processing Horn and the other hand to fully tighten the Converter s connection by locking the metal wrench to the Converter s wrench grooves Flow Tubing Assembly There are two holes in the Sample Processing Horn for the purpose of circulating water You will need to determine an appropriate length for both sides and then cut the provided Flow Tubing into two If y2. To Select or Modify a Program or Sequence Press HERE op Options Touch a Selection OO S00 S00 EPISONIC PAGE 9 Memory Menu Select Modify a Program allows you to create or modify up to 10 different programs for sonication at specific user selected time intervals Select Modify Sequence 1 or 2 allows you to select a se quence of programs to run in successive order There is a limit of 6 programs in one sequence Options allows you to select Celsius or Fahrenheit settings enable footswitch operation and adjust the LCD screen s display contrast Program Menu EpiSonic can store up to 10 programs in the memory To select or load a program press a number on the keypad on the screen the selected program number will display above the Amplitude box After touching each of the following fields the background will change to yellow to indicate the selected field Use the keypad to enter values and then press ENT Enter to com plete the entry Press CLR Clear to erase any values Amplitude sets the desired starting amplitude Process Time sets the desired total sonication time or total Pulse ON Time Pulse OFF Time is not counted Pulse ON Time ON Cycle sets the desired sonication time of one duty cycle before a Pulse OFF cycle begins Leaving this blank will run the sonication continuously e SelectiModify a Program e SelectiModify SEQUENCE 1 e SelectiMo
3. I TOUCHSCREEN CONTROL The EpiSonic Multi functional Bioprocessor is controlled through a touchscreen liquid crystal display LCD user interface After in stalling and plugging in the instrument you can turn on the power switch at the rear panel of the sonication generator Main Menu Manual Operation allows you to start and stop sonication manually and adjust amplitude settings in real time Program Menu allows you to create and run a program with presets and store it in the system memory or create and run a sequence of programs Options allows you to select Celsius or Fahrenheit settings enable footswitch operation and adjust the LCD screen s display contrast Manual Operation Amplitude adjust the intensity of the sonication energy by touching the UP or DOWN arrows Power shows the current power output of ultrasonic en ergy in Watts Energy shows the accumulated amount of ultrasonic en ergy transmitted in Joules Elapsed Time shows the total time of actual sonication Probe Temperature if the Temperature Probe accessory is installed this shows the current temperature of the water Start Pause press START to begin sonication which can be followed by STOP pressing START again will reset all values Press PAUSE to pause sonication which can be followed by RESUME this will not reset your values USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 up To Operate Manually Press HERE
4. on Page 7 Do not use any lubricants grease or WD 40 on any of the parts unless instructed otherwise For maximum protection this preventive maintenance should be performed weekly assuming regular use 2 Clean The Sampling Processing Horn The top surface of the sample processing horn should be gently cleaned with a mild soap solution and wiped down with a wet cloth or towel Do not use any cleaning solvents or abrasives Do not let any water drip into the converter Remember to drain any water after use The sample processing horn is constructed from aluminum and you will notice a bit of oxidation over time This is normal and it has no effect on the performance of the sample processing horn 3 Inspect Connections and Cables Inspect the converter cable and power cord to ensure there is no fraying or damage to the rubber sleeve or insulation Never use a cable with broken end connections exposed wires or frayed insulation USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 17 ACCESSORIES I COMPATIBLE COOLING SYSTEMS A cooling system is not necessary in most cases for DNA and chromatin shearing If you intend to use EpiSonic for extended periods of time hard to process samples or industrial use you can obtain the following cooling systems for use with EpiSonic PERISTALTIC PUMPS Watson Marlow PumpPro Watson Marlow PumpPro Economy Manostat Carter Multichannel Pump CHILLERS amp RECIRCULATI
5. COM TABLE OF CONTENTS PrECAUHONS 44554632000 054 545004004 40t Geese Bee ewe eee Sne 4 Safety PrecautionS c6 crac heed nee ese cutee ee ohn FURR EEE SRE OST RB Oe RN RES 4 Equipment Protection s ws warn a tee ea wee wee ee EE 4 INTTOGUCHON EE 5 Background Information os6 4 04 oe0cee vas bathe od hid ee owns es eeoe ao bod teens 5 Technical Specifications 0 0 eee eee teen eee eee ees 5 COMDONGIGS 644 cc ease ec LEENE e agen ee eee see ENGSTE FEEN 6 Pals Te te e EE 6 Rear Panel Diagram neve vad od eae ane doh oe eos deo abe es hee es Gan eae es 6 JS dE o BEE 7 Pre Assembly abs w oe sus eww a 6 Geb OO DO GEESE AE Oba eae Pena Re EN eae RY 7 M in ASSeMDIy 6 4440454 cosas cone danseneneeneeee ae een edurdacigesseeyenecn 8 SelLUD Pigs I 1 4 vacuo sone beeen reese mee eee be REER 8 ODEON EEN 9 Touchscreen Control EE 9 Handling Your SampleS EE 13 Optimization Suggestions sueueueue eee saben 4 hee aeRO Reed eS bee es 14 Example Protocols cc ccc cece reece c eee eee e eee eneeees 16 Pre Optimized DNA Shearing 0 0 cc eee eee eee eee ees 16 Pre Optimized Chromatin Shearing 0 0 cc ccc eee eee eee eee 16 Caring For Your Device 1 ccc tc tc tc tc ee eee reece eee eeaee 17 After Operation 17 Maintenance EE 17 PCCOSSOVICS d i E bode E pGr eene E eee eee soe 18 Compatible Cooling Systems 18 EpiSonic ACCESOMIGS 26 so vGewe done ee Ge bw dR eee eae 2 h6 Re RS ee bee bees 18 USA
6. ON and Pulse OFF times com bined Sequence Menu After creating programs in the Program Screen you can link several programs together to run in succession You can save up to two different sequences After touching each of the following fields the background will change to yellow to indicate the selected field Use the keypad to enter values and then press ENT Enter to com plete the entry Press CLR Clear to erase any values NUM represents the position in the succession in which the corresponding program will run PGM the program field will allow you to enter the desired program that will run for that NUM position Unused PGM fields should have a dash in the box Delay After allows you to set a delay or rest time after the corresponding program has been run in the sequence Save Run press SAVE if you want to store the program in the memory Press RUN to continue to the Program Opera tion screen USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 PROGRAM H Amplitude Process WOR LO ep eeh 00 00 00 Pulse OHW WIRT I d Time 00 00 10 Pulse OFF 00 00 10 Power Km W Time ey EE Program Progress M o GE we d de P EXIT SEQUENCE SEQUENCE Trt ROG NUM PGM Delay After P enn _00 00 00 J 2 2 00 00 00 ii 00 00 00 4 5 6 NN if ry Q i d di d ENT 00 00 00 00 00 00 00 00 00 EPISONIC PAGE 11 Sequence Operation T
7. TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 3 PRECAUTIONS The EpiSonic Multi Functional Bioprocessor has been designed constructed and tested to assure your safety as the operator of the sonication device However for complete safety and protection of this equipment PLEASE READ ALL OF THE INSTRUCTIONS IN THIS MANUAL CAREFULLY PRIOR TO OPERATION INCLUDING THE FOLLOWING WARNINGS I SAFETY PRECAUTIONS m Do NOT open the sonication generator cover without supervision by an authorized service provider m Do NOT open the ultrasonic converter or high voltage cables without supervision by an authorized service provider m Do NOT operate the sonication generator without any connection or sound processing horn attached to the high voltage cable as this may pose a shock hazard m Do NOT attempt to disconnect the high voltage cable while the sonication generator is running as this may pose a shock hazard m Do NOT plug in the power cord on either side while the sonication generator s power switch is on m Do NOT touch any open cable connections on the sonication generator while the power is on m Do NOT immerse the ultrasonic converter in liquids of any sort or let condensed moisture or liquid drip into the ultrasonic con verter m Do NOT touch the activated sample processing horn or ultrasonic converter as it may cause burns or tissue damage m Do NOT operate the sonication generator while the soundproof encl
8. CE Marked Evaluated to UL 61010 1 2nd Ed IEC 61010 1 CAN CSA C22 2 No 61010 1 2nd Ed IEC 61010 1 Mod WEEE Directive on Waste Electrical and Electronic Equipment This product contains electrical or electronic materials which must be disposed of mess properly The end user should dispose this product in an environmentally friendly way through a separate collection and not as unsorted waste For more information consult your institution s disposal guidelines or e mail info epigentek com USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 5 vil IL multi functional bioprocessor COMPONENTS I PARTS FOR ASSEMBLY Sonication Generator Soundproof Enclosure Sample Processing Horn Ultrasonic Converter ee Ki Includes strap wrench and spanner wrench for assembling these two parts together AC Power Cord Flow Tubing Converter Cable ll REAR PANEL DIAGRAM Converter Power Cable Port Cord Port a CONVERTOR On Off she gaan 120 VAC 50 60 Hz 7A MAX Switch Cooling Fan W A Brooklyn NY 11226 U S A FOR 100 120 VA SE BA FAST A TING FUSE MODEL EQC 1000 e ht Foot Switch CE deet DO NOT REMOVE COVE Jack TEMPERATURE PROBE FOOTSWr USB Port A S_57Q08A 05 10 emperature for software Probe Port updates PAGE 6 EPISONIC WWW EPIGENTEK COM INFO EPIGENTEK COM INSTALLATION Prior to installing your EpiSonic device please take note of the following
9. Cat EQC 1000 ATP Cat EQC 1000 AFS Cat EQC 1000 X38 Cat EQC 1000 X96 Cat EQC 1000 X02 Cat EQC 1000 X05 Cat EQC 1000 X15 Cat P 2025 48 48 rxn P 2025 96 96 rxn Cat P 2026 48 48 rxn P 2026 96 96 rxn Cat EQC 1000 PSG Cat EQC 1000 PPH Cat EQC 1000 PSE Cat EQC 1000 PUC Cat EQC 1000 PPC Cat EQC 1000 PFT Cat EQC 1000 PCC WWW EPIGENTEK COM INFO EPIGENTEK COM This page is left intentionally blank USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 19 E PI TE EN TEK Copyright 2011 Epigentek Group Inc All Rights Reserved Complete Solutions for Epigenetics EPISONIC is a trademark of Epigentek Group Inc
10. Erlo Ie multi functional bioprocessor USER MANUAL 1000 sertes gt DNA SHEARING A gt CHROMATIN SHEARING Pm CELL DISRUPTION Fee e GEES SECHS gt HOMOGENIZATION gt AND MORE Manufactured in the USA Epigentek Group Inc _ a EPI fs EN TEK Web www epigentek com E mail info epigentek com Complete Solutions for Epigenetics Tel 1 877 374 4368 USA Tel 1 718 484 3990 nt Fax 1 718 484 3956 PAGE 2 EPISONIC WARRANTY INFORMATION Epigentek warrants that the EPISONIC 1000 sonication generator device is free from defects in material and workmanship under normal use and service for twenty 20 months from the date that the warranty period begins unless otherwise indicated Warranty periods begin on the date of delivery All warranties apply only to the original purchaser of this product Epigentek s obligation under this warranty is limited to the repair or replacement at its sole discretion of any product or part thereof if the product is deemed to have a manufacturing defect upon examination by Epigentek or its authorized repair service provider The sample processing horn is covered under a 6 six month warranty from defects in material and workmanship under normal use and service Included soundproof enclosure box cables tubing and tools are not covered by any warranties Discoloration or paint wear is not covered by any warranties Any warranties shall cover only manufacturing defects a
11. ON SYSTEMS Model 3006 capacity 6L Model WK 230 Benchtop Chiller capacity 6L Oasis 160 Compact Recirculating Chiller Thermo Fisher Thermo Fisher Thermo Fisher Thermo Fisher Thermo Fisher Solid State Cooling Systems Cat 14 283 2 Cat 14 283 3 Cat 13 875 241 Cat 13 874 7 Cat 22 037 157 Cat HOASIS160 DISCLAIMER The rate of water circulation may become an additional variable in your optimization procedures The flow tubing can be fed through the bottom of the soundproof enclosure However you may need to obtain new flow tubing or tubing connectors usually Tygon tubing to make sure it can fit into your cooling system Consult the manufacturer of the system for further information Epigen tek is not responsible for damage to or malfunction with the EpiSonic device from improper cooling system connections ll EPISONIC ACCESSORIES APPROVED ACCESSORIES Temperature Probe Footswitch 384 Well PCR Plate 96 Well PCR Plate 0 2 ml PCR Vial 0 5 ml PCR Vial 1 5 ml Microtube COMPATIBLE KITS ChromaFlash One Step ChIP Kit ChromaFlash One Step Magnetic ChIP Kit REPLACEMENT PARTS Sonication Generator Sample Processing Horn Soundproof Enclosure Ultrasonic Converter AC Power Cord Flow Tubing Converter Cable PNG T8 EPSON ies Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek Epigentek
12. d easily Other samples may require higher sonication intensity or a longer processing duration Due to variability between each type of sample you process you will need to optimize the sonication conditions for maximum efficiency To minimize variability in results use the following recommended steps towards optimizing your conditions 1 Use a Proper Vessel The most suitable sample vessels for optimal transmission of sonication energy are composed of polystyrene or polycarbonate Poly propylene sample vessels are also suitable but since the material can absorb some of the sonication energy before it reaches the sample extended sonication times or higher intensity may be required Avoid using brittle plastics which may crack when exposed to long periods of ultrasonic energy See Handling Your Samples on Page 13 for examples of suggested vessels 2 Maximize Energy Contact For the best transmission of sonication energy into your sample vessel ensure that the bottom of the vessel touches the surface of the sample processing horn When filling the horn with water try to make sure the vessel does not float away from the surface The amount of water added should be just above the sample level so that the sample is completely submerged Avoid overflowing or add ing the water past the vessel s height 2 Adjust Amplitude Power The higher the amplitude setting the more energy power output will be transmitted to your sample and the quicker t
13. dify SEQUENCE 2 e Options Touch a Selection TOWEN 2 Humber Key for Another Memory 2 3 Process Time Pulse OH Time 4 J E Pulse OFF Time 7 H t Temperature Monitoring PROGRAM SETTINGS PROGRAM 0 Amplitude A d n CLR C ENT louch a Parameter Box ert P to Change the Value RUN E JI Pulse OFF Time OFF Cycle sets the desired wait time of one duty cycle after a Pulse OFF cycle ends Leaving this blank will also run the sonication continuously Note that one Pulse ON cycle and one Pulse OFF cycle equals one duty cycle Temperature Monitoring activates the Temperature Probe accessory if installed Shutdown Temperature if Temperature Monitoring is active you can set a temperature at which the program will pause if that tem perature is reached You will be able to resume the sonication when the temperature drops below that value Save Run press SAVE if you want to store the program in the memory Press RUN to continue to the Program Operation screen PAGE 0 EPSON ies WWW EPIGENTEK COM INFO EPIGENTEK COM Program Operation This is similar to the Manual Operation screen However it will also display the Process Time Pulse ON Time Pulse OFF Time and Shutdown Temperature settings programmed in the previous Program Menu Probe Temperature cannot be toggled while the program is in progress Program Progress shows the progress of the current pro gram It includes both Pulse
14. he sample will be processed However higher amplitude will heat up the water in the sample processing horn more quickly Keep in mind that even with the same amplitude setting the power output can vary if you change the ves sel size type and or composition in addition to the amount of water added Therefore when changing these variables in such a case you will need to adjust the amplitude ac cordingly as the sonication runs to reach the desired power output but you will want to minimize the amount of adjusting needed by starting as close as possible to the correct amplitude Use the following chart as a reference point before starting the sonication process Then while the sonication runs adjust the amplitude until the power output is f Note This chart is only a general guideline and reached It is always better to set the amplitude at a lower setting and to adjust upwards was tested with a 96 well PCR plate 3 Maintain Water Temperature When the sonication energy is being generated it will also generate heat which will thus increase the temperature in your water and sample You will want to avoid degradation of your sample or reduction in processing efficiency due to overheating For DNA and chromatin shearing maintain the water temperature as cool as possible but not to be lower than 4 C or exceed 37 C unless specifi cally instructed in an Epigentek approved protocol for use with EpiSonic For extended sonication periods y
15. his is similar to the Program Operation screen It will display the current program being run in the sequence as indicated on the top left above Amplitude Program Progress shows the progress of the current pro gram It includes both Pulse ON and Pulse OFF times com bined Sequence Progress shows the progress of the current se quence It includes both Pulse ON and Pulse OFF times of all programs combined in addition to any delay intervals in between each program Options Menu Temperature Units if you have the Temperature Probe ac cessory installed you can select whether to display the tem perature readings in Celsius or Fahrenheit Footswitch Operation if you have the Footswitch accessory installed you can choose the desired method of manual op eration with the Footswitch Display Contrast adjust the LCD screen s contrast by making it lighter left or darker right PNG 2 EPRISONTCI SEQU 1 PROGRAM W Amplitude A Set 00 00 00 Pulse OH 00 00 20 TAU H all gt sl Time j ut Hi Pulse OFF 8 107 nna Time 00 00 d ach i a E Temperature coer o M Program Progress i o SEQUENCE Progress eme 0 TART 9 l i REV 41 31 16 Temperature Units D Celsius Fahrenheit i Footswitch Operation Press and hold for Sonication Release to stop Press once for Sonication Press again to stop i DISPLAY CONTRAST fa _ 4 _____ D Changes Saved Automa
16. ill allow sonication energy to transmit better You can sonicate the water in the sample processing horn for several minutes to degas the water 8 Maintain Sample Consistency While optimizing your sonication procedure you must maintain the following constant sample conditions a sample volume b sample concentration c sample type or source d cross linking fixation duration for ChIP e concentration of formaldehyde for ChIP use less than 1 and f antibody lot if applicable 9 Monitor and Record Parameters Make sure to keep a record of your sonication parameters in order to achieve reproducibility or to compare against different param eters Note the following a power output energy delivered b amplitude setting c sonication ON and OFF times d total sonica tion duration e water temperature f rate of water circulation if applicable g when ice was added or water level was adjusted if applicable h sample vessel used i conditions of the assay sample itself and j any other parameters that you believe may affect processing results Additional Tips for Chromatin Shearing In addition to the above optimization suggestions the following are also important when working with chromatin e Cell Tissue Fixation Appropriate chromatin cross linking is required Insufficient or over cross linking will cause DNA loss or in creased background Make sure the cross linking time 10 15 min concentration of formaldeh
17. it at least three minutes before reapplying power Make sure your electrical outlet has proper grounding before plugging in the sonication generator Only use parts provided or approved by Epigentek for use with your EpiSonic device Install your EpiSonic device according to this manual s instructions Failure to follow the instructions including hand tightening the sample processing horn and converter instead of using the included wrenches may result in damage of the instrument PAGE 4 EPISONIC WWW EPIGENTEK COM INFO EPIGENTEK COM INTRODUCTION Congratulations on obtaining your new EpiSonic Multi Functional Bioprocessor from Epigentek You made the right choice by select ing EpiSonic as your primary means for processing your biological samples The sample processing horn is designed for high through put processing with 96 well or 384 well plates but is also suitable for single sample processing if desired The innovative design of the sample processing horn allows for the transmission of an equal distribution of sonication energy throughout the entire processing surface leading to a higher consistency and reproducibility in results than other waterbath based methods and b less harsher pro cessing and less over fragmentation than probe based methods I BACKGROUND INFORMATION With the EpiSonic Multi Functional Bioprocessor AC line power is transformed into a 20 KHz signal which is then converted to a mecha
18. nd do not cover any damage resulting from accidents misuse abuse neglect modifications repair or service by any unauthorized parties exposure to extreme temperatures exposure to acids or chemical sol vents failure to comply with the instructions in this user manual or maintenance not in accordance with any information contained in literature accompanying the product Any unauthorized modifications repair or servicing by parties not approved by Epigentek shall void any warranties All returns for repairs or replacement must be accompanied with a Return Merchandise Authorization number RMA which can be obtained through Epigentek s customer service department Equipment should be returned in the original packaging If you no longer have the original packaging you will need to contact Epigentek for approved packaging methods Damage occured from return ship ping due to poor packing is not covered by this warranty WHEN RETURNING ANY MATERIALS TO EPIGENTEK YOU MUST CERTIFY THAT THE MATERIALS BEING RETURNED ARE FREE OF ANY HAZARDOUS INFECTIOUS OR RADIOACTIVE SUBSTANCES AND ALL PARTS HAVE BEEN PROPERLY DECONTAMINATED EPIGENTEK GROUP INC EpiSonic Multi Functional Bioprocessor 1000 Made in the United States of America USA Phone 1 877 374 4368 International Phone 1 718 484 3990 Ordering E mail sales epigentek com Support E mail support epigentek com Website http www epigentek com WWW EPIGENTEK COM INFO EPIGENTEK
19. nical vibration due to the characteristics of internal piezoelectric crystals The vibration is amplified and transmitted through the sample processing horn in ultrasonic waves which longitudinally expands and contracts The distance the waves travel is dependent on the amplitude selected by the operator in the touchscreen menu Thus as the setting for the amplitude increases so will the sonication intensity within the sample being processed When the ultrasonic waves travel through liquid by rapid vibration of the sample processing horn the formation of microscopic bub bles called cavitation begins These cavitation bubbles will continuously absorb energy until they implode This primary mechanism of sample processing releases an immense energy in the cavitation field to disperse or break up biomolecules perfect for also fragment ing DNA or chromatin as a result ll TECHNICAL SPECIFICATIONS 13 5 Ibs 6 1 kgs 19 5 L x 11 W x 5 25 H 49 5 cm x 27 9 cm x 13 3 cm 24 5 Ibs 11 1 kgs 10 L x 10 W x 16 5 H 25 5 cm x 25 5 cm x 42 cm 1000 V rms max 20 KHz nom Operating Environment Temp 39 95 F 4 35 C Relative Humidity 20 90 Non Condensing Indoor Use Only Shipping Environment Temp 35 120 F 2 49 C Relative Humidity 10 90 Non Condensing Ambient Pressure Extremes 40 000 ft 12 000 meters Compliancy Restriction of Hazardous Substances ROHS Compliant Directive 2002 95 EC Regulatory Information
20. osure box is open or this may result in inury to your ears m Inspect high voltage cable for cracks or rips in the protective outer jacket Do NOT operate the sonication generator with a dam aged cable or this may result in serious injury m Do NOT use any substitute or third party cables only use approved cables specifically designed for use with EpiSonic Power off and unplug the sonication generator from both sides before attempting to replace any fuses This device is intended for use in research applications and has not been tested or authorized for diagnostic use Il EQUIPMENT PROTECTION m Do NOT place the EpiSonic device in direct sunlight or near magnets Install the device in an area free from dirt excessive dust and explosive or corrosive fumes Keep the device away from extreme temperature and humidity Do NOT place the generator within a fume hood gw Do NOT allow the vibrating horn to touch any hard surfaces or it may overload the sonication generator become damaged or cause damage to the surface m Do NOT turn off the sonication generator while it is in the middle of sonication Press STOP on the touchscreen menu prior to disabling power m Do NOT let water sit for extended periods of time in the sonication processing horn if unused If the sample processing horn is attached to the converter do NOT hold this assembly by the converter as this may bend or damage the threaded connection In the event of loss of power wa
21. ou can maintain water temperature by adding ice chips note that this will alter your water level as the ice melts by changing the water completely by using a peristaltic pump or by using a recirculating chiller see page 18 for Accessories For an exact temperature reading we highly recom mend the EpiSonic Temperature Probe accessory Epigentek Cat EQC 1000 ATP 5 Adjust Sonication Timing Because every sample is different along with variations in the sample vessel temperature and water level it is important to determine appropriate pulse on and off program durations and cycles Sonication durations will vary based upon different sample sources and sample concentrations used Every sample is different some are harder to shear or process than others Controlling the pulse on and off durations will also help regulate the rate of which the temperature increases 6 Test Vessel Rotation Although the EpiSonic 1000 is designed to evenly distribute sonication energy via the entire surface of the sample processing horn it may be beneficial to rotate the vessel for more difficult to shear samples As processing times will vary you will need to determine your rotation pattern at even intervals e g 20 seconds ON then turn the microplate 90 degrees during the 20 seconds OFF WWW EPIGENTEK COM INFO EPIGENTEK COM 7 Test Degasification For difficult to process samples degassed water removal of carbon dioxide and or oxygen w
22. ou do not intend to install a cool ing system an exact length is not as crucial Ensure there is a pinch clamp on both tubings and firmly connect the end of each tubing to the Sample Processing Horn s holes There are also two holes at the bottom of the Soundproof En closure for the Flow Tubing to pass through Custom length Tygon tub ing can also be obtained at your own discretion USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 7 ll MAIN ASSEMBLY Connect one end of the Converter Cable to the bottom of the Converter Make sure it is securely attached by turning the fastening mechanism clockwise at the end of the cable Open the lid of the Soundproof Enclosure and feed the oth er end of the Converter Cable through the opening at the bottom until you can rest the Converter comfortably and firmly in the opening Connect and fasten the other end of the Converter Cable to the rear panel of the Sonication Generator Secure the fastening mechanism lil SETUP DIAGRAM Soundproof Enclosure Sonication Generator Power Cord gt l ee Converter Cable PAGE 8 EPISONIC Connect the Power Cord to the rear panel of the Sonica tion Generator Plug the Power Cord into a grounded pow er outlet The device is now installed and ready for use Sample Processing Horn Converter Flow Tubing With Pinch Clamps WWW EPIGENTEK COM INFO EPIGENTEK COM OPERATION
23. stic rack to Use a plastic rack to Description to be processed in to be processed in hold one or multi hold one or multi hold one or multi each PCR well Seal each PCR well Seal ple vials containing ple vials containing ple vials containing with well caps or with well caps or samples Make sure samples Make sure samples Make sure adhesive film adhesive film the vial is capped tbe vial is capped the vial is capped properly prior to properly prior to properly prior to sonication sonication sonication Minimum 4 ul well 5 ul well 5 ul well 10 ul well 50 ul well Sample Volume Maximum 15 ul well 60 ul well 60 ul well 150 ul well 400 ul well Sample Volume Suggested Volume 10 ul well 30 ul well 30 ul well GO ul well 200 ul well Suggested Sample 0 01 to 0 02 ug per 0 01 to 0 02 ug per 0 01 to 0 02 ug per 0 01 to 0 02 ug per 0 01 to 0 02 ug per DNA Concentration ul of buffer ul of buffer ul of buffer ul of buffer ul of buffer Suggested Sample Chro 0 1 to 0 2 ug per ul 0 1 to0 2 ug perul 0 1 to0 2 ug perul 0 1 to 0 2 ug per ul 0 05 to 0 1 ug per matin Concentration of buffer of buffer of buffer of buffer ul of buffer Epigentek EQC 1000 X38 EQC 1000 X96 EQC 1000 X02 EQC 1000 X05 EQC 1000 X15 Catalog Number USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 13 PAGE UE TEP S O mes Ili OPTIMIZATION SUGGESTIONS Certain samples will shear or process quite fast an
24. tically on EXIT EAI d WWW EPIGENTEK COM INFO EPIGENTEK COM ll HANDLING YOUR SAMPLES The proper sample vessel is critical to obtaining optimal and reproducible results All sample vessels should be surrounded by water on all sides so there is an even distribution of sonication energy Do not use traditional closed flat bottomed microplates since water will not be able to wrap around each well The bottom of the vessel should touch or be as close as possible to the surface of the sample processing horn for maximum expo sure to ultrasonic energy m Avoid using vessels made of thick or hardened plastic Non brittle thin walled polystyrene and polycarbonate material is sug gested for optimal sonication otherwise non brittle thin polypropylene plastic can be used m The actual sample inside your vessel should be completely submerged just below the water level Be careful not to cross contaminate wells when handling vial well caps or adhesive covering film Wear gloves m If desired rotate your sample vessel while processing at consistent intervals to see if this helps to optimize your conditions Use the following table to help determine which vessel is most appropriate for your samples Vessel Vessel Type 384 Well PCR Plate 96 Well PCR Plate 0 2 mi PCR Vial 0 5 ml PCR Vial 1 5 ml Microtube Samples Processed 1 384 1 96 1 96 1 42 1 20 ina Single Run Handling Add your samples Add your samples Use a plastic rack to Use a pla
25. w times to siphon the water from the sample processing horn into the beaker or sink make sure the beaker or sink is below the height of the sample processing horn Remember to close the pinch clamps afterwards 3 Dry the Sample Processing Horn Using a clean cloth or paper towel thoroughly wipe down any excess water or moisture inside and around the sample processing horn and the soundproof enclosure Do not leave unused water in the sample processing horn for extended periods of time as this may damage the surface Be careful not to get water or moisture on the ultrasonic conductor ll MAINTENANCE The design of EpiSonic allows for easy self maintenance Regular maintenance is necessary to facilitate optimal performance It is recommended to periodically inspect the unit both visually and physically Before performing any maintenance procedure make sure the power switch is in the OFF position with the power cord unplugged from the outlet 1 Clean and Tighten Connective Areas Sonication devices create intense vibration which can put stress on or loosen the threaded connection between the converter and the sample processing horn over time The converter and sampling processing horn should be disassembled with the wrench set The threading hole and flat surfaces around this area should be cleaned with a cotton swab dipped in isoproyl alcohol Let the parts air dry and reassemble tightly with the wrench set as instructed in Pre Assembly
26. yde 1 as final concentration and quanch solution 0 125 M glycine are correctly used during the cross linking step of chromatin preparation e Chromatin Quality Make sure that chromatin is properly extracted from a sufficient amount of cells or tissues Chromatin extract should be properly stored at 80 C USA TEL 1 877 374 4368 INT TEL 1 718 484 3990 EPISONIC PAGE 15 EXAMPLE PROTOCOLS I PRE OPTIMIZED DNA SHEARING Please contact an Epigentek representative for specific protocol information ll PRE OPTIMIZED CHROMATIN SHEARING Please contact an Epigentek representative for specific protocol information PAGE I EP SON ies WWW EPIGENTEK COM INFO EPIGENTEK COM CARING FOR YOUR DEVICE I AFTER OPERATION When you are finished processing your samples and do not intend to continue to use the EpiSonic again within a few hours you will need to perform the following steps 1 Power Off the Unit Make sure the sonication program is no longer processing or running PAUSE or STOP You can now safely turn the power switch off at the rear panel of the sonication generator Do not leave the EpiSonic on and unattended for extended periods of time as this may damage the LCD touchscreen display 2 Drain Out the Water Simply open the soundproof enclosure lid and take one of the flow tubings and place the end in a beaker or sink Open the pinch clamp on the tubing and pinch the tubing with your fingers a fe
Download Pdf Manuals
Related Search