InviMag Blood DNA Maxi Kit/ IG User manual
Contents
1. A suitable to loaded No Assay Selected v reagents Version 1 Reagen loading Jisp tip loading Status Ready m Se service Figure 7 Assay Selection screen of the InviGenius software Select the assay file DLBD E400 S2000 and ensure that the field suitable to loaded reagents is checked in order to list only assay files that are compatible with the loaded reagents If no assay is visible in the list there is either an error related to reagent loading blocked barcode label or at least one reagent container has been loaded that was already in use and does not contain enough reagent to run with the selected amount of loaded samples If the assay selection was successful proceed with disposable tip loading Disposable Tip Loading Disposable tips loading 1 Tip Load Position 3 LLLA Em al Disp sheaths Assay selection loading Status Ready User service Figure 8 Disposable tip loading screen There are three tip rack positions available in the InviGenius system Fig 8 A1 A3 that can be loaded with 1100 ul conductive filtered tips to guarantee efficient prevention of cross contaminations of samples and reagents Remaining tip numbers in the rack are shown in field B Tip numbers can be changed by pressing the number field B and manually enter the number of available tips Empty tip racks can be unloaded and reloaded by 15 InviMag Blood2. stratecee molecular User manual InviMag Blood DNA Maxi Kit IG for use onthe InviGenius STRATEC Molecular GmbH for automated purification of DNA from 2 ml whole blood samples with magnetic beads 2431320x00 aak STRATEC Molecular GmbH D 13125 Berlin Fe Instructions for InviMag Blood DNA Maxi Kit IG The InviMag Blood DNA Maxi Kit IG combines the advantages of the innovative Invisorb technology with easy handling of magnetic particles in combination with the InviGenius robotic platform for an efficient and reliable isolation of nucleic acids with a high purity The InviMag Blood DNA Maxi Kit IG is the ideal tool for walk away automated isolation and purification of highly pure total genomic and mitochondrial DNA from 2 ml whole blood samples stabilized with EDTA or citrate but not heparin The kit is designed for use on the InviGenius workstation The interplay of the DNA extraction and purification chemistry provided by the InviMag Blood DNA Maxi Kit IG was intensely tested and validated The DNA binding magnetic particles are characterized by a high surface area uniform size distribution and good suspension stability and therefore are highly suitable for high throughput processing Due to the high purity the isolated DNA is ready to use in a broad panel of downstream applications or can alternatively be stored at 20 C for subsequent use The kit is neither validated for the isolation of genomi
3. 4 ml dd H O mix thoroughly and store diluted and unused Proteinase K at 20 C Add 36 ml of 96 100 ethanol to the bottle Wash Buffer Il Mix thoroughly and always keep the bottle firmly closed Add 110 ml of 99 796 isopropanol to the bottle Binding Solution and always keep the bottle firmly closed Reagents and equipment to be supplied by user Measuring cylinder 250 ml Pipette tips Disposable gloves ddH O Vortexer 96 100 ethanol 99 7 Isopropanol O0o00000 InviMag Blood DNA Maxi Kit IG 0414 The InviMag Blood DNA Maxi Kit IG is validated with 2 Propanol Rotipuran gt 99 7 p a ACS ISO Order no 6752 from Carl Roth Possible suppliers for Isopropanol Carl Roth Applichem 2 Propanol 2 Propanol f r die Molekularbiologie Rotipuran gt 99 7 p a ACS ISO Order no A3928 Order no 6752 Important indications 1 Minimum volume of samples in primary tubes Sigma 2 Propanol Order no 59304 1L F The procedure of the InviMag Blood DNA Maxi Kit IG has been optimized for the isolation of genomic DNA from up to 2 ml human whole blood We advise to provide at least 2 5 ml blood per sample tube 12 mm diameter more if a wider diameter is used to prevent pipetting distribution errors during processing and to avoid unwanted flagged sample results due to used low sample volumes 2 Sample volume smaller than 2 ml For samples with a volume s 2 ml please adjust the final volume with ei
4. cross contaminations and reagent cross overs are effectively eliminated by the automated purification process The use of unique bar codes for samples and reagents prevents unwanted transpositions The InviGenius instrument uses magnetic rods to transport the DNA binding magnetic particles through the various extraction phases such as binding washing and elution The volume of reagents required for DNA isolation is reduced to a minimum Eliminating the direct liquid handling and increasing the automation level results in a fast reliable and robust technique After a sample specific lysis using Lysis Buffer HLT and Proteinase K optimal binding conditions are adjusted by addition of Binding Solution The genomic DNA binds to the simultaneously added magnetic particles and is separated from solution using magnetic rods which are controlled by the InviGenius system Subsequent to seven washing steps of the particle bound nucleic acids pure DNA is finally eluted Due to the high purity the eluted total DNA genomic and mitochondrial is ready to use in a broad panel of downstream applications PCR real time PCR Restriction enzyme digestion HLA typing o Southern Blotting OO O0 For the isolation of DNA from single blood samples STRATEC Molecular offers the Invisorb Spin Blood Mini Kit or for 8 96 samples the Invisorb DNA Blood Mini HTS 96 Kits for use on a centrifuge vacuum manifold or other robotic stations For further inform
5. defined as a range from 15 30 C Quality Control and product warranty STRATEC Molecular warrants the correct function of the InviMag Blood DNA Maxi Kit IG for applications as described in this manual Purchaser must determine the suitability of the product for its particular use Should any product fail to perform the applications as described in the manual STRATEC Molecular will check the lot and if STRATEC Molecular investigates a problem in the lot the product will be replaced free of charge STRATEC Molecular reserves the right to change alter or modify any product to enhance its performance and design at any time In accordance with STRATEC Molecular s ISO 9001 2000 and ISO EN 13485 certified Quality Management System the performance of all components of the InviMag Blood DNA Maxi Kit IG have been tested separately against predetermined specifications routinely on lot to lot to ensure consistent product quality If you have any questions or problems regarding any aspects of InviMag Blood DNA Maxi Kit IG or other STRATEC Molecular products please do not hesitate to contact us A copy of STRATEC Molecular s terms and conditions can be obtained upon request or are presented at the STRATEC Molecular webpage For technical support or further information please contact from Germany 49 0 30 9489 2901 2910 from abroad 49 0 30 9489 2903 2907 or contact your local distributor InviMag Blood DNA Maxi Kit IG 0414 Intend
6. firmly closed Resuspend a lyophillized Proteinase K tube with 2 4 ml dd H5O and mix thoroughly Add 36 ml of 96 100 ethanol to the bottle Wash Buffer mix thoroughly and always keep unused bottles firmly closed One run is defined as a run using 12 samples Please keep in mind that partially used bottles may be used in other runs The InviGenius tracks the sample numbers and runs that were extracted with each bottle MAP Solution B and Proteinase K may be only used in two runs for quality mixing InviMag Blood DNA Maxi Kit IG 0414 Symbols Manufacturer Lot number Catalogue number Date of manufacture Expiry date Consult operating instructions Temperature limitation Do not reuse 9x HKE 8 SE Storage All buffers and kit contents of the InviMag Blood DNA Maxi Kit IG except Proteinase K and MAP Solution B magnetic beads should be stored at room temperature and are stable for at least 12 months at this condition Proteinase K IG Lyophilized Proteinase K should be stored at 2 8 C Dissolved Proteinase K must be stored at 20 C MAP Solution B IG The magnetic beads should be stored at 2 8 C Wash Buffer and Binding Buffer Wash Buffer and Binding Buffer charged with either ethanol or isopropanol should be stored at room temperature and must be appropriately sealed If there are any precipitates visible within the provided solutions solve them by carefully warming up to 0 C Room temperature RT is
7. high temperature or UV irradiation Careful isolation and handling of high molecular weight DNA is necessary to ensure its functionality in various downstream applications Damaged DNA could perform poorly in applications such as genomic Southern blotting long template PCR and construction of cosmid libraries Handling fresh and stored material before the extraction of DNA For the isolation of genomic DNA use either fresh or frozen samples 20 C This procedure minimizes degradation of crude DNA by limiting the activity of endogenous nucleases Storage of DNA Store the genomic DNA at 2 8 C or 20 C Keep in mind that the storage of genomic DNA at 20 C may cause shearing particularly if the DNA is exposed to repeated freezing and thawing cycles Plasmid DNA and other small circular DNAs can be stored at 2 8 C or at 20 C without risk 22 InviMag Blood DNA Maxi Kit IG 04 14 Troubleshooting Problem Pipetting distribution errors Low concentration of extracted DNA Degraded or sheared DNA No assay selectable Eluted DNA is brownish colored Probable cause Comments and suggestions Transfer of Proteinase K failed Ensure that lyophilized Proteinase K is resuspended with the appropriate volume of water Samples transfer failed The sample tube must contain at least 2 5 ml sample Reagent buffer transfer failed Ensure that the Wash Buffer and Binding Buffer incomplete is filled up properly with ethanol isopropan
8. 0 InviMag Blood DNA Giga Kit IG 8 x 12 preps 2431320400 Invisorb Spin Blood Mini Kit 50 preparations 1031100200 Invisorb Spin Blood Mini Kit 250 preparations 1031100300 Invisorb Spin Blood Midi Kit 50 preparations 1031110300 Invisorb Spin Blood Midi Kit 250 preparations 1031110500 InviGenius and consumables InviGenius 1 unit 5011100000 Starting Box I IG 1 box 2400110100 Sheath Box Conductive filter tips 1 ml 2 x 2 rack pack 384 pieces 5 Waste Trays 120 sample tubes Sheath Bundle 10 x 48 pieces 5011100300 Sheaths 1000 pieces 5011100200 Conductive filter tips 1 ml 10 x 96 pieces 5011100400 Waste tray IG 25 pieces 5011100100 Possible suppliers for Isopropanol Carl Roth Applichem Sigma 2 Propanol 2 Propanol f r die Molekularbiologie 2 Propanol Rotipuran gt 99 7 p a ACS ISO Order no A3928 Order no 59304 1L F Order no 6752 24 InviMag Blood DNA Maxi Kit IG 04 14 HEU ELICIT LORRI ILLE LIQUET CUT ILLE HT stratecee molecular STRATEC Molecular GmbH Robert R ssle Str 10 13125 Berlin Germany Phone 49 30 94 89 29 01 Fax 49 30 94 89 29 09 E mail info berlin 2 stratec com www stratec com 1G3a03 IG 04 2014
9. 1 late E Free lanas amp LHAp Svea loadiney Management Status Ready User service Figure 10 Plate loading screen In general an Incubation Plate D Nerbe Plus 12 well reservoir DWP is used at the incubator position a Working Plate A 2 ml Sarstedt DWP is used at the working position and an Elution Plate E SLG stripes is used at the elution position Plates can be unloaded reloaded by 1 Pressing the plate position directly A The software will focus the plate position on the main screen 2 Pressing the Unload button B 3 The plate can be reloaded by pressing on the offered plate s in C Attention Be aware that it is not possible to manually enter used rows of a plate within the system software If a partially filled plate is removed from within the system software all used lanes of that plate will be deleted from the system memory One run with the DNA Blood Maxi Kit on the InviGenius using 12 samples will require one new Incubation Plate D at the incubator position one new Working Plate A at the working position and one free row within the Elution Plate E located at the eluate position Please ensure that the depicted lanes displayed on the monitor are consistent with the real lanes in the corresponding positions To avoid contaminations never wash or reuse consumed plates 17 InviMag Blood DNA Maxi Kit IG 04 14 Waste management Ensure that the waste tray capacity is sufficient
10. 12 InviMag Blood DNA Maxi Kit IG 04 14 Sample Loading After Loading was selected the sample loading screen is shown Select Samples A to proceed with the sample loading process Loading m ng Despasahbim shnzhs PAL ET ET TD TA T wg Micraglatieg Waste BA TOR ere Status Ready Figure 4 Loading submenu of the InviGenius software Samples loading Usar service o SAMPLE 1 edited SAMPLE 2 edited o SAMPLE 3 edited 2 SAMPLE 4 edited SAMPLE 5 edited e SAMPLE 6 edited Id SAMPLE 7 edited SAMPLE 8 edited SAMPLE 9 edited SAMPLE 10 edited c SAMPLE 11 edited SAMPLE 12 edited d Figure 5 Sample loading screen of the InviGenius software Prepare the sample rack with primary sample tubes that are prefilled with blood from which the DNA shall be extracted Sample tubes are not provided within the kit The operator must either use primary tubes provided from the blood collection blood bank etc In case that no primary tubes are available common collection tubes can be ordered at Sarstedt for example order no 55 476 5 ml tubes 75x12 mm polystyrene For each reaction a total sample volume of 2 ml is processed However we advise that the total provided sample volume should be at least 2 5 ml to ensure stable processing If primary tubes are used that are completely filled with blood 2 5 ml please premix by inverting the tube before usage to ens
11. DNA Maxi Kit IG 04 14 1 Press the Loading Position directly The software will focus the loading position on the main screen 2 Press the Unload Button C 3 The loading position can then be refilled with a new tip rack by pressing on the corresponding tip rack and then selecting the tip type D Note Disposable tips are not supplied within the kit We recommend the use of validated conductive tips which can be ordered at STRATEC Molecular Offered tips are 50 pl conductive tips 10x 96 pieces order no 5011120100 and 1100 ul conductive tips 10x 96 pieces order no 5011120200 Ensure that conductive tips are used otherwise the liquid level detection LLD will not work which will lead to an abort of the run Due to the installed tip allocation module of the pipettor it is required that only filter tips are used Non filter tips will be rejected by the system To avoid contaminations never wash or reuse consumed tips Disposable Sheaths Loading The sheaths function as protection devices for the magnetic rods and are picked up automatically during the run Disposable sheaths loading Sheath Load Position G80808080808 4 002028080808 lanes CCLI 020808080808 to Sheath Type Dap Sp toading Status Ready Es Uzer service Figure 9 Disposable sheaths loading screen The loading procedure of the disposable sheaths works analogous to the disposable tip loading screen For each run always 12 di
12. InviMag Blood DNA Maxi Kit IG 0414 Safety information When and while working with chemicals always wear a suitable lab coat disposable gloves and protective goggles Avoid skin contact Adhere to the legal requirements for working with biological material For more information please consult the appropriate material safety data sheets MSDS These are available online in convenient and compact PDF format at www stratec com for each STRATEC Molecular Product and its components If buffer bottles are damaged or leaking WEAR GLOVES AND PROTECTIVE GOGGLES when discarding the bottles in order to avoid any injuries STRATEC Molecular has not tested the waste generated by the InviMag Blood DNA Maxi Kit IG procedures for residual infectious materials Contaminations of the waste with residual infectious materials is highly unlikely but cannot be excluded completely Therefore all generated waste has to be considered infectious and should be handled and discarded accordingly to local safety regulations Below European Community risk and safety phrases for the components of the InviMag Blood DNA Maxi Kit IG are listed Lysis Buffer HLT Wash Buffer HLT Proteinase K contains guanidine hydrochloride H302 315 319 P280 305 351 338 H315 319 334 335 P280 305 351 338 310 405 H315 Causes skin irritation H319 Causes serious eye irritation H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled H335 May ca
13. and elimination of ethanol Elution of pure DNA After lysis the DNA binds to the magnetic beads whereas contaminations and enzyme inhibitors are efficiently removed during the following seven washing steps At the end highly purified DNA is eluted Oo O O Q Lysis Samples are lysed in a Deep Well Reservoir Plate at elevated temperatures in the presence of Lysis Buffer HLT and Proteinase K Binding After addition of Binding Solution and MAP Solution B magnetic beads to the lysate the DNA is bound to the beads Removing residual contaminants Contaminants are efficiently removed using Wash Buffer HLT and Wash Buffer while the DNA remains bound to the magnetic beads Elution The DNA is finally eluted in Elution Buffer M The eluted DNA is ready to use for different subsequent downstream applications like PCR amplification digestion with restriction enzymes Southern hybridizations HLA typing etc InviMag Blood DNA Maxi Kit IG 0414 Yield and quality of genomic DNA The amount of purified DNA using the InviMag Blood DNA Maxi Kit IG procedure from whole blood depends on the leucocytes content sample source transport condition storage and age Typically a volume of a 2 ml whole blood sample from a healthy individual with an elevated white blood cell content ranging from 3x10 to 1x10 cells per ml will yield at least 20 ug of genomic DNA The overall purified DNA yield is about 20 50 ug DNA derived from 2 ml
14. ation please contact 49 0 30 9489 2901 2910 in Germany 49 0 30 9489 2903 2907 from foreign countries or ask your local distributor InviMag Blood DNA Maxi Kit IG 0414 Sampling and storage of starting material For reproducible and high yields the appropriate sample storage is essential Yields may be varying from sample to sample depending on factors such as health of the donor sample age sample type transport and storage conditions Blood Best results will be obtained with fresh whole blood samples Mammalian blood samples stabilized with EDTA or citrate but not heparin are stable at room temperature 18 25 C for 2 3 hours after collection For short term storage up to 24 h samples can be stored at 4 8 C For long term storage freezing the samples at 20 C or 80 C is recommended Avoid multiple thawing and freezing cycles of samples before isolating the DNA because this may lead to degradation of the DNA Various different primary tubes blood collection systems e g Sarstedt Greiner and anticoagulants except heparin can be used to collect blood samples for the InviMag Blood DNA Maxi Kit IG procedure STRATEC Molecular will not take responsibility if other sample types are used than described Principle and procedure The InviMag Blood DNA Maxi Kit IG procedure comprises the following steps Lysis of blood cells and protein digestion Binding of DNA to magnetic beads Washing of the bead bound DNA
15. c DNA from cell cultures tissues blood cards dried blood stains nor urine The application of the kit for isolation and purification of viral DNA has neither been tested nor evaluated For research use only Trademarks InviMag Invisorb InviGenius Registered marks trademarks etc used in this document even when not specifically marked as such are not to be considered unprotected by law The Invisorb technology is covered by patents and patent applications US 6 110363 US 6 043 354 US 6 037 465 EP 0880535 WO 9728171 WO 9534569 EP 0765335 DE 19506887 DE 10041825 2 WO 0034463 InviMag Invisorb InviGenius are registered trademarks of STRATEC Biomedical AG The PCR process is covered by US Patents 4 683 195 and 4 683 202 and foreign equivalents owned by Hoffmann La Roche AG 2014 STRATEC Molecular all rights reserved InviMag Blood DNA Maxi Kit IG 0414 Table of Content Kit contents of the InviMag Blood DNA Maxi Kit IG Symbols Fehler Textmarke nicht definiert Storage Fehler Textmarke nicht definiert Quality Control Fehler Textmarke nicht definiert Intended use Product use limitation Safety information Product characteristics of the InviMag Blood DNA Maxi Kit IG Sampling and storage of starting material Principle and procedure Yield and quality of genomic DNA Important notes Preparing reagents and buffers Reagents and equipment to be supplied by user Important indications Gen
16. ed use The InviMag Blood DNA Maxi Kit IG is designed for fully automated extraction and purification of genomic DNA from up to 2 ml whole blood in the range of 1 12 blood samples using the InviGenius robotic platform The nucleic acid isolation protocol is suitable for routinely walk away automated preparation of DNA from fresh or frozen blood samples For reproducible and high yields an appropriate sample storage is essential see Sampling and storage of the starting material page 8 Common blood collection tubes not provided and anticoagulants EDTA and citrate but not heparin can be used All utilities reagents and plastics besides filter tips required for preparation of DNA from blood are provided by the InviMag Blood DNA Maxi Kit IG THE PRODUCT IS INTENDED FOR USE BY PROFESSIONALS SUCH AS TECHNICIANS PHYSICIANS AND BIOLOGISTS TRAINED IN MOLECULAR BIOLOGICAL TECHNIQUES It is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of DNA RNA followed by signal detection or amplification Any diagnostic results generated by using the sample preparation procedure in conjunction with any downstream diagnostic assay should be interpreted with regard to other clinical or laboratory findings To minimize irregularities in diagnostic results adequate controls for downstream applications should be used Product use limitation The kit is only validated for the ext
17. eral overview of the InviGenius system Preparing and loading of the InviGenius system PREPARING THE REAGENTS PREPARING THE SYSTEM SAMPLE LOADING REAGENT LOADING ASSAY SELECTION DISPOSABLE TIP LOADING DISPOSABLE SHEATHS LOADING PLATE LOADING WASTE MANAGEMENT BATCH DEFINITION BATCH CHECKING BATCH PROCESSING AFTER A RUN DAILY MAINTENANCE UV DECONTAMINATION FEHLER TEXTMARKE NICHT DEFINIERT General notes on handling DNA Troubleshooting Ordering Information O SSO OC qe O0 0 qi 22 23 24 InviMag Blood DNA Maxi Kit IG 0414 Kit contents of the InviMag Blood DNA Maxi Kit IG Store the MAP Solution B at 2 8 C Store lyophilized Proteinase K at 2 8 C Dissolved Proteinase K must be stored at 20 C Store all other kit components at room temperature RT Catalogue No 2431320100 HERREN Lysis Buffer HLT 4 x 80 ml 2 runs per bottle 8 x 80 mg proteinase edo 8 x 2 4 ml working solution Binding Solution 4 x empty bottles a fill with 99 796 Isopropanol final volume 4x 110 ml runs per bottle 4 x 36 ml Wash Buffer final volume 4x 120 ml 2 runs per bottle Elution Buffer M 4 x 60 ml 2 runs per bottle 1 run per plate Incubation Plate D Elution Plate E 1 8 runs per plate Sheath Box 2 x 48 sheaths racks 4 runs per rack Initial steps Add 110 ml Isopropanol molecular biologic grade undenaturated to each empty bottle labeled with Binding Solution Keep unused bottles
18. for the planned assay If not empty the solid waste container Handle the generated waste accordingly as required by local disposal laws Waste management Waste capacity 190 Dropshalstatus In place Fill level 0 disposables 0 Number of wasted disposable sheaths 0 Number of wasted disposable tips 0 Figure 11 Waste management screen After the waste container was emptied click on Empty solid waste to reset the filling level status Batch definition Batch definition Assay descriptions MES to loaded reac MES DBLD 400 S2000 Version 1 Aliquots Volume 2000 2000 Wasa maa garmer Status Ready es Se service Bmch checking Figure 12 Batch definition screen This screen allows de selection of the desired assay and sample numbers in one informational screen The assay can be switched by using the two arrow buttons A If samples have to be excluded from the batch use the arrows B to switch between samples and click on the Remove from batch button field B If deselected samples have to be added to the process list the button Remove from batch will be changed to Add to batch By default all loaded and recognized samples are selected 18 InviMag Blood DNA Maxi Kit IG 04 14 Batch checking This screen shows a summary of all loaded disposables samples and buffers in one screen and functions as a final security check In case of any error the problem po
19. m User interaction with the InviGenius instrument is performed by use of the touch LCD J located at the top front right side of the instrument 11 InviMag Blood DNA Maxi Kit IG 04 14 Preparing and loading of the InviGenius system Preparing the reagents Before starting a run prepare one vial of Proteinase K and add ethanol to the Wash Buffer and isopropanol to the Binding Solution bottle as indicated in Tab 1 see page 3 or page 9 Preparing the system Turn on the InviGenius system using the power switch located at the back right side of the instrument The InviGenius software will automatically be loaded after the system has booted up The door of the InviGenius system must be closed during system initialization and runs After booting and successful initialization of the InviGenius system a log in screen appears Figure 2 Log in with the provided user name and password Login User identifier Password Status Ready am Figure 2 Log in screen of the InviGenius software After logged in the main screen of the InviGenius software is displayed Figure 3 Select Loading A to proceed with loading of the system or select Processing B to define and run an assay if the system has been loaded already Main Menu Cate managamari b Mainienance Pasminisiratian InviGenius Status Ready User service Figure 3 Main menu of the InviGenius software
20. nce Periodical maintenance Cleanup 7 Diagnostics procedures ss Cleanup Recovery drop caicher Status Ready m Ler service Figure 16 Maintenance screen of the InviGenius software In the UV decontamination menu adjust the exposure time A and finally press the Start button B During the decontamination process the instrument door will be locked to prevent any UV radiation release in the lab Warning UV radiation is harmful It causes serious burns of the skin and leads to irreparable damage of the eyes and skin Ensure that no lab personnel is submitted to direct UV light Do not try to force open the instrument door during the decontamination process UV decontamination A Duration 00 15 00 I HH MM SS Remaining tme 00 00 00 UV decontaminaton status There was no UV decontamination B completed tS E Figure 17 UV decontamination screen When the decontamination is finished go back to the main menu by using the Back button The device is now decontaminated and can be either switched off or used for sample processing We recommend decontaminating the instrument daily 21 InviMag Blood DNA Maxi Kit IG 04 14 General notes on handling DNA Nature of DNA The length and delicate physical nature of DNA requires careful handling to avoid damage due to shearing and or enzymatic degradation Other conditions that affect the integrity and stability of DNA include acidic and alkaline environments
21. of this screen Reagent Loading The reagent loading process is analogous to the sample loading procedure Reagents loading z Barcode Name Exp date Unknown Unknown PG410055130900441505 Elution B M PG 2015 05 31 PG618034301302931505 MAP Sol B PG 2015 05 31 PG306108131800771505 Wash B II PG 2015 05 31 PG706024340102531505 Proteinase K PG 2015 05 31 PG218098100000621505 Binding Sol PG 2015 05 31 Unknown Unknown a PG124078130700521505 Lysis B HLT PG 2015 05 31 Unknown Unknown WE sample loading Hack Status Ready OO o service AALALA AAA AAA TTY Figure 6 Reagent loading screen of the InviGenius software Decap all reagents before inserting them into the reagent rack Ensure that the bar code labels face to the right side of the loading bay The order of inserted reagents with the same bottle size is not important because the type and position of each reagent is identified by the unique reagent bar code and the bar code at the corresponding loading position of the reagent rack However the possible loading positions are limited by the size of useable bottles After rack insertion the loading status of the reagents is shown In case of unsuccessful reagent identification remove the rack check the bar code orientation and repeat the loading process slowly 14 InviMag Blood DNA Maxi Kit IG 04 14 Assay Selection Assay selection Assay description DBLD E400 S2000
22. ol Do not reuse bottles more often than described in Tab 1 Blood components settled In case of large sample volumes 222 ml carefully premix the sample tube before inserting it into the sample rack No too much ethanol Assure that the Wash Buffer Binding Buffer is isopropanol added to Wash filled up with ethanol isopropanol properly as Buffer Binding Buffer indicated in Tab 1 Incorrect storage of starting Ensure that the storage condition of the starting material material was correct Avoid multiple freezing and thawing cycles of the sample Old material Ensure that the starting material is fresh or stored at appropriate conditions for long term storage at 20 C Old material often contains degraded DNA Combination of reagents from Assure that only reagents belonging to one kit type different kits or blocked barcode are used a combination of reagents belonging to during reagent loading procedure different kit types is not supported by the system Ensure that the reagent barcode label is visible within the reagent rack window Small part of the magnetic Centrifuge the eluates at full speed for 1 min and particles are left in the elution transfer supernatant to a new plate tube 23 InviMag Blood DNA Maxi Kit IG 04 14 Ordering information Product Package size Catalogue No InviMag Blood DNA Maxi Kit IG 8 x 12 preps 2431320100 Related products InviMag Blood DNA Mini Kit IG 8 x 12 preps 243112010
23. plication Note The run duration of the Blood Maxi DNA assay using 12 samples is approximately 200 minutes After a run After the run is completed and no additional run should be started remove all plates and reagents from the instrument and store them accordingly to GLP guidelines Please keep in mind that the plates could contain possibly infectious material Important Never erase not completely filled plates from within the software if the plates shall be used for another run It is not possible to register partially used plates within the software An erased plate will lead to loss of all stored information As with all medical clinical and diagnostically equipment all waste products liquids tips sheaths and plates should be treated as potentially dangerous bio hazard waste Daily maintenance UV decontamination The InviGenius system is equipped with an internal UV lamp 254 nm wavelength that should be used daily either at the end of the working day or in the morning before a run is started The suggested decontamination time is about 20 min To start the UV decontamination go to the main menu of the InviGenius software and select Maintenance Main Menu Process na C InviGenius Logout Status Ready a HU m Ier developer Figure 15 Main screen of the InviGenius software When the sub item Maintenance is opened select UV decontamination 20 InviMag Blood DNA Maxi Kit IG 04 14 Maintena
24. raction of human whole blood samples The isolation of DNA from other sources like stool samples tissues bacteria fungi or viruses was neither tested nor validated No guarantee in operability is issued if other samples than whole blood are used The included chemicals are only useable once Differing of starting material may lead to inoperability Therefore neither a warranty nor guarantee in this case will be given implied or expressed The user is responsible to validate the performance of the STRATEC Molecular product for any particular use STRATEC Molecular does not provide validations of performance characteristics of the product with respect to specific applications STRATEC Molecular products may be used e g in clinical diagnostic laboratory systems conditioned upon the complete diagnostic system of the laboratory the laboratory has been validated pursuant to CLIA 88 regulations in the U S or equivalents in other countries All products sold by STRATEC Molecular are subject to extensive quality control procedures according to ISO 9001 2000 and ISO EN 13485 and are warranted to perform as described herein Any problems incidents or defects shall be reported to STRATEC Molecular immediately upon detection thereof The chemicals and the plastics are for laboratory use only They must be stored in the laboratory and must not be used for other purposes than intended The product with its contents is not suitable for consumption
25. sition is highlighted in red color To clear a displayed error click directly on the red highlighted field and follow the instructions printed on the instrument screen If no error is displayed proceed by pressing the button Batch processing Batch checking Selected assay DBLD E400 2000 Sach defmitiorn Hack Batch processing Status Ready User service Figure 13 Batch definition screen Batch processing After the instrument door has been closed the run can be started by pressing the Start Button A The door will be locked during the complete run The door will only be unlocked after a run has been successfully finished or an error occur that demands user interaction Do not try to force open the door during a run or the run will be aborted If there is an error the Start button is blocked The user will not be able to start the instrument until the error has been solved To do so go back to Batch definition and solve the error see chapter Batch definition for detailed information Batch processing Batch identifier Assay description Actual step s Remaining me 00 00 00 Process state Idle Events p pus 4 Status Ready a L Se service Figure 14 Batch processing screen 19 InviMag Blood DNA Maxi Kit IG 04 14 At the end of the process the nucleic acid containing eluates are located in the appropriate eluate position and can be used for any further downstream ap
26. sposable sheaths one row of the sheaths rack are picked up regardless of the sample number processed This is done to assure that all rods are protected against contaminations at all time In general the number of sheaths supplied within the kit is sufficient for the amount of runs imprinted on the kit package In case of low or not enough available sheaths a reorder at STRATEC Molecular 100 pieces bulk order nr 5011120300 or 10x 48 pieces order no 5011120400 is possible Comparable to the disposable tips loading procedure the number of rows left in the tip rack can be defined by pressing on the number area Please ensure that the disposable sheaths are loaded and displayed consistent to the manually entered sheaths rows in the rack to guarantee correct sheaths pick up Don t remove single disposable sheaths within a row of the sheaths rack if less than 12 samples are processed within one run There is a sheaths detection sensor installed in the instrument If less than 12 sheaths are picked an error message is displayed and all picked up sheaths will be discarded into the waste container before the next row of sheaths are picked up for verification To avoid contaminations never wash or reuse consumed sheaths 16 InviMag Blood DNA Maxi Kit IG 04 14 Plate Loading Analogous to the previous loading screens the incubation working and elution plates are loaded within the plate loading screen Figure 10 oplates Fraa lanas
27. ther distilled water or 1x PBS to 2 5 ml 3 Elution volume The final processed elution volume is 400 ul After the elution step an aliquot of about 380 yl is transferred to the elution plate Typically 400 ul eluate will contain about 20 50 ug of purified DNA 10 InviMag Blood DNA Maxi Kit IG 04 14 General overview of the InviGenius system Figure 1 Layout of the InviGenius system There are three plate positions available in the InviGenius system which can be loaded with the corresponding plates the incubation A working B and elution position C The lysis is performed at the incubation position A whereas the washing and elution process is performed at the working position B The eluate containing the extracted nucleic acids will be finally transferred to the elution position C There are three loading positions for disposable tip trays D1 D3 and one position E for the disposable sheaths available The loading bay F is located at the very right side of the instrument The sample rack is loaded into the far left lane whereas the reagent rack is located at the right position of the loading bay The moveable Magnetic Separation Head MSH G is located on top of the incubator lid parking position The automatic pipettor head H is installed above the loading bay parking position The disposable waste tray I is located behind the lower cover of the InviGenius syste
28. ure a homogenous solution Always decap inserted sample tubes Keep in mind that only 12 positions of the sample rack can be processed per run due to the limited number of wells per row of the plastic ware For correct identification of the sample tubes bar codes if available must face to the bar code scanner window located at the right side of the loading bay 13 InviMag Blood DNA Maxi Kit IG 04 14 After inserting the sample rack in the very left lane of the loading bay an updated screen will show the identifiers read out from the sample bar codes Figure 5 In case of unsuccessful sample identification remove the rack check the bar code orientation and reinsert the rack slowly If no sample barcodes is present the corresponding sample will be flagged as unknown sample In that case the sample name has to be entered changed manually using the Edit button Keep in mind that samples marked as unknown sample will not be processed It is possible to rename barcode recognized samples too Select the corresponding sample using the arrow buttons followed by the Edit button and enter the desired sample name After a short time period about 5 min the bar code scanner will be deactivated In that case the operator has to restart the scanner by pressing the START SCANNER button After all samples have been loaded recognized and or renamed proceed with reagent loading by selecting Reagent loading on the bottom right hand side
29. use respiratory irritation P280 Wear protective gloves protective clothing eye protection face protection P305 P351 P338 If in eyes Rinse cautiously with water for several minutes Remove contact lenses and continue rinsing P310 Immediately call a POISON CENTER or doctor physician P405 Store locked up Emergency medical information can be obtained 24 hours a day from infotrac outside of USA 1 352 323 3500 inside of USA 1 800 535 5053 InviMag Blood DNA Maxi Kit IG 0414 Product characteristics of the InviMag Blood DNA Maxi Kit IG The InviMag Blood DNA Maxi Kit IG is the ideal tool for an efficient and fully automated DNA extraction and purification from fresh or frozen whole blood samples using magnetic beads in combination with the InviGenius robotic platform preparation 20 50 ug depending on the blood sample about 170 min Az60 A280 transport storage age 12 samples 1 7 1 9 and source 2 ml fresh or frozen whole blood stabilized with EDTA or citrate but not heparin The DNA isolation process is based on the interaction of nucleic acids with silica coated magnetic particles at adapted buffer conditions The InviGenius instrument will automatically perform the whole DNA purification procedure including sample and reagent distribution without any user intervention except the initial loading of the system This allows safe handling of potentially infectious samples whereas
30. whole blood However if a whole blood sample is mixed with anticoagulant containing buffer solutions the overall leukocyte concentration decreases and the yield of the DNA extraction procedure is reduced slightly Important notes Important points before starting a protocol After receiving the kit check all components for visible damage If buffer bottles are damaged or leaking contact the STRATEC Molecular Technical Services or your local distributor In case of liquid spillage refer to Safety information page 6 Do not use damaged kit components because their use may lead to poor kit performance o When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles Discard gloves if they become contaminated Do not combine components of different kits Avoid microbial contamination of the kit reagents To minimize the risk of diseases from potentially infectious material we recommend working under laminar air flow for all steps performed outside the InviGenius platform o This kit should only be used by trained personnel OOo0oo0 Preparing reagents and buffers Before starting a run equilibrate all reagents at room temperature Where necessary gently mix and redissolve any precipitates by incubating at 30 C Swirl gently to avoid foaming Lysis Buffer HLT and Elution Buffer M are ready to use 8 x 12 DNA extractions Resuspend a lyophilized Proteinase K tube by addition of 2
Download Pdf Manuals
Related Search