NucleoMag® 96 Tissue - MACHEREY
Contents
1. Genomic DNA from tissue User manual NucleoMag 96 Tissue May 2014 Rev 03 MACHEREY NAGEL www mn net com Genomic DNA from tissue Table of contents 1 Components 1 1 Kit contents 1 2 Equipment and consumables to be supplied by user 5 2 Product description 6 2 1 The basic principle 6 2 2 Kit specifications 6 2 3 Magnetic separation systems 7 2 4 Adjusting the shaker settings 7 2 5 Handling of beads 8 2 6 Elution procedures 9 3 Storage conditions and preparation of working solutions 10 4 Safety instructions 11 5 Protocol for the isolation of genomic DNA from tissue 13 6 Appendix 19 6 1 Troubleshooting 19 6 2 Ordering information 21 6 3 Product use restriction warranty 22 MACHEREY NAGEL 05 2014 Rev 03 3 Genomic DNA from tissue 1 Components 1 1 Kit contents NucleoMag 96 Tissue 1x 96 preps 4 x 96 preps 24 x 96 preps REF 744300 1 744300 4 744300 24 NucleoMag B Beads 2x 1 5mL 12 mL 70 mL Lysis Buffer T1 50 mL 100 mL 1000 mL Binding Buffer MB2 45 mL 180 mL 2x500 mL Wash Buffer MB3 75 mL 300 mL 2 x 900 mL Wash Buffer MB4 75 mL 300 mL 2 x 900 mL Wash Buffer MB5 125 mL 500 mL 3 x 1000 mL Elution Buffer MB6 30 mL 125 mL 2 x 500 mL Proteinase K lyophilized 75 mg 4x75 mg 24x75 mg Proteinase Buffer PB 8 mL 15 mL 3x 35 mL User manual 1 1 1 For preparation of working solutions and storage conditions see section 3 4 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissu2. 225 uL of cleared lysate to a Square well Block for further processing 5 600 x g 5 min 225 uL cleared lysate 3 Bind DNA to NucleoMag B Beads 24 uL NucleoMag B Beads 360 pL MB2 Mix by shaking for 5 min at RT Optional Mix by pipetting up and down Remove supernatant after 2 min separation MACHEREY NAGEL 05 2014 Rev 03 13 NucleoMag 96 Tissue Wash with MB3 Remove Square well Block from NucleoMag SEP 600 pL MB3 Resuspend Shake 5 min at RT Optional Mix by pipetting up and down Remove supernatant after 2 min separation Wash with MB4 Remove Square well Block from NucleoMag SEP 600 pL MB4 Resuspend Shake 5 min at RT Optional Mix by pipetting up and down Remove supernatant after 2 min separation o gt Wash with MB5 Leave Square well Block on NucleoMag SEP 900 pL MB5 Incubate for 45 60 s Note Do not resuspend the beads in Buffer MB5 Remove supernatant 14 MACHEREY NAGEL 05 2014 Rev 03 NucleoMag 96 Tissue 7 Elute DNA Remove Square well Block from NucleoMag SEP 50 200 pL MB6 Optional Elute at 55 C Shake 5 min at RT Optional Mix by pipetting up and down Separate 2 min and transfer DNA into elution plate tubes MACHEREY NAGEL 05 2014 Rev 03 15 NucleoMag 96 Tissue Detailed protocol This protoc
3. any and all damages resulting from such application DNA RNA PROTEIN purification products of MACHEREY NAGEL are suitable for N VITRO USES ONLY ONLY MACHEREY NAGEL products specially labeled as IVD are also suitable for IN VITRO diagnostic use Please pay attention to the package of the product IN VITRO diagnostic products are expressly marked as IVD on the packaging IF THERE IS NO IVD SIGN THE PRODUCT SHALL NOT BE SUITABLE FOR IN VITRO DIAGNOSTIC USE ALL OTHER PRODUCTS NOT LABELED AS IVD ARE NOT SUITED FOR ANY CLINICAL USE INCLUDING BUT NOT LIMITED TO DIAGNOSTIC THERAPEUTIC AND OR PROGNOSTIC USE No claim or representations is intended for its use to identify any specific organism or for clinical use included but not limited to diagnostic prognostic therapeutic or blood banking It is rather in the responsibility of the user or in any case of resale of the products in the responsibility of the reseller to inspect and assure the use of the DNA RNA protein purification products of MACHEREY NAGEL for a well defined and specific application MACHEREY NAGEL shall only be responsible for the product specifications and the performance range of MN products according to the specifications of in house quality control product documentation and marketing material This MACHEREY NAGEL product is shipped with documentation stating specifications and other technical information MACHEREY NAGEL warrants to meet the stated specifica
4. by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnet Remove and discard supernatant by pipetting MACHEREY NAGEL 05 2014 Rev 03 17 NucleoMag 96 Tissue B mE Wash with MB5 Leave the Square well Block on the NucleoMag SEP magnetic separator Note Supernatant is colorless magnetic bead pellet is clearly visible Gently add 900 uL Buffer MB5 to each well and incubate for 45 60 s while the beads are still attracted to magnets Then aspirate and discard the supernatant Note Do not resuspend the beads in Wash Buffer MB5 This step is to remove traces of ethanol and eliminates a drying step Elution Remove the Square well Block from the NucleoMag SEP magnetic separator Add desired volume of Buffer MB6 50 200 uL to each well of the Square well Block and resuspend the beads by shaking 5 10 min at 56 C Alternatively resuspend beads completely by repeated pipetting up and down and incubate for 5 10 min at 56 C Separate the magnetic beads by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnets Transfer the supernatant containing the purified genomic DNA to the Elution Plate Note Yield can be increased by 15 20 by using pre warmed elution buffer 55 C or by incubating the bead elution buffer suspensio
5. distance of the separation plate from the magnetic pins and the volume to be processed The individual times for complete attraction of the beads to the magnetic pins should be checked and adjusted on each system It is recommended using the separation plates or tubes specified by the supplier of the magnetic separator Washing the beads Washing the beads can be achieved by shaking or mixing In contrast to mixing by pipetting up and down mixing by shaker or magnetic mixing allows simultaneous mixing of all samples This reduces the time and number of tips needed for the preparation Resuspension by pipetting up and down however is more efficient than mixing by a shaker or magnetic mix 8 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue Method Resuspension Number of tips efficiency needed Magnetic mix Low Shaker Low Pipetting High acceptable good excellent 8 channel pipetting device 2 6 Elution procedures Purified DNA can be eluted directly with the supplied Elution Buffer MB6 Elution can be carried out in a volume of 50 UL It is essential to cover the NucleoMag Beads completely with elution buffer during the elution step The volume of dispensed elution buffer depends on the magnetic separation system e g the position of the pellet inside the separation plate For efficient elution the magnetic bead pellet should be resuspended completely in the elut
6. min For optimal lysis mix occasionally during incubation Make sure that the lysis tubes are securely closed If RNA free DNA is crucial for downstream applications an RNase digest may be performed add 20 uL RNase A 20 mg mL solution not included see ordering information and incubate for additional 5 min at room temperature Clear lysates Centrifuge the samples for 5 min at a full speed 5 600 6 000 x g Remove cap strips Transfer 225 uL of the cleared lysate equilibrated to room temperature to a Square well Block Do not moisten the rims of the well Note See recommendations for suitable plates or tubes and compatible magnetic separators section 1 2 16 MACHEREY NAGEL 05 2014 Rev 03 NucleoMag 96 Tissue Bind DNA to NucleoMag B Beads Add 24 uL of NucleoMag B Beads and 360 uL Buffer MB2 to each well of the Square well Block Mix by pipetting up and down 6 times and shake for 5 min at room temperature Alternatively when processing the kit without a shaker pipette up and down 10 times and incubate for 5 min at room temperature Note NucleoMag B Beads and Buffer MB2 can be premixed before use Premix just before use storage of premixed beads and buffers is not recommended Mix 24 uL NucleoMag B Beads with 360 uL Buffer B2 per sample Depending on the dead volume of the reservoir additional amounts of bead suspension and binding buffer are necessary Use 384 uL of the suspension per we
7. or a rash occurs Get medical advice attention Bei Hautreizung oder ausschlag Arztlichen Rat einholen rztliche Hilfe hinzuziehen If experiencing respiratory symptoms Call a POISON CENTERF doctor Bei Symptomen der Atemwege GIFTINFORMATIONSZENTRUM Arzt anrufen Get medical advice attention Bei anhaltender Augenreizung rztliche Rat einholen rztliche Hilfe hinzuziehen Store in a well ventilated place Keep container tightly closed Beh lter dicht verschlossen an einem gut bel fteten Ort aufbewahren Store in a well ventilated place Keep cool Beh lter dicht verschlossen an einem gut bel fteten Ort aufbewahren For further information please see Material Safety Data Sheets www mn net com Weiterf hrende Informationen finden Sie in den Sicherheitsdatenbl ttern www mn net com 12 MACHEREY NAGEL 05 2014 Rev 03 NucleoMag 96 Tissue 5 Protocol for the isolation of genomic DNA from tissue Protocol at a glance For additional equipment and hardware requirements refer to section 1 2 and 2 3 respectively For detailed information on each step see page 17 Before starting the preparation Check if Proteinase K was prepared according to section 3 1 Lyse samples up to 20 mg tissue up to 1 x 10 cells or bacteria pellet from up to1 mL overnight culture 25 uL Proteinase K 200 uL T1 Mix 56 C 1 3 h or overnight 2 Clear lysates by centrifugation transfer
8. 61 P 280 P 301 312 P 302 352 P 304 340 P 305 351 313 P 312 P 330 P 333 313 P 342 311 P 337 313 P 403 233 P 403 235 Keep away from heat hot surfaces sparks open flames and other ignition sources No smoking Von Hitze hei en Oberfl chen Funken offenen Flammen sowie anderen Z ndquellenarten fernhalten Nicht rauchen Keep container tightly closed Beh lter dicht verschlossen halten Avoid breathing dust Einatmen von Staub vermeiden Wear protective gloves eye protection Schutzhandschuhe Augenschutz tragen IF SWALLOWED Call a POISON CENTER doctor if you feel unwell BEI VERSCHLUCKEN Bei Unwohlsein GIFTINFORMATIONSZENTRUM Arzt anrufen IF ON SKIN Wash with plenty of water BEI KONTAKT MIT DER HAUT Mit viel Wasser waschen IF INHALED Remove victim to fresh air and keep at rest in a position com fortable for breathing BEI EINATMEN An die frische Luft bringen und in einer Position ruhigstellen die das Atmen erleichtert IF IN EYES Rinse continuously with water for several minutes Remove contact lenses if present and easy to do continue rinsing BEI KONTAKT MIT DEN AUGEN Einige Minuten lang behutsam mit Wasser sp len Vorhandene Kontaktlinsen nach M glichkeit entfernen Weiter aussp len Call a POISON CENTER doctor if you feel unwell Bei Unwohlsein GIFTINFORMATIONSZENTRUM Arzt anrufen Rinse mouth Mund aussp len IF skin irritation
9. EY NAGEL GmbH amp Co KG Tel 49 24 21 969 270 tech bio mn net com Trademarks KingFisher is a registered trademark of Thermo Fisher Scientific NucleoMag is a registered trademark of MACHEREY NAGEL GmbH amp Co KG Te MagS is a trademark of Tecan Group Ltd Switzerland All used names and denotations can be brands trademarks or registered labels of their respective owner also if they are not special denotation To mention products and brands is only a kind of information i e it does not offend against trademarks and brands and can not be seen as a kind of recommendation or assessment Regarding these products or services we can not grant any guarantees regarding selection efficiency or operation MACHEREY NAGEL 05 2014 Rev 03 23
10. ads Buffer MB2 and the NucleoMag B Beads are added to the lysate After magnetic separation the paramagnetic beads are washed twice to remove contaminants and salts using Wash Buffer MB3 and Wash Buffer MB4 There is no need for a drying step as ethanol from previous wash steps is removed by a final incubation of the beads in Buffer MB5 Finally highly purified DNAis eluted with low salt elution buffer Buffer MB6 and can directly be used for downstream applications The NucleoMag 96 Tissue kit can be used either manually or automated on standard liquid handling instruments or automated magnetic separators 2 2 Kit specifications NucleoMag 96 Tissue is designed for rapid manual and automated small scale preparation of highly pure genomic DNA from tissue samples cells or bacteria using the NucleoMag SEP see ordering information or other magnetic separation systems see section 2 3 Manual time for the preparation of 96 samples is about 120 minutes The purified DNA can be used directly as template for PCR blotting or any kind of enzymatic reactions NucleoMag 96 Tissue allows easy automation on common liquid handling instruments The actual processing time depends on the configuration of the instrument and the magnetic separation system used Typically 96 samples can be purified in less than 120 minutes using the NucleoMag SEP on the automation platform The kit provides reagents for the purification of up to 20 ug of pure gen
11. anty contract tort including negligence or strict liability arising in connection with the sale or the failure of MACHEREY NAGEL products to perform in accordance with the stated specifications This warranty is exclusive and MACHEREY NAGEL makes no other warranty expressed or implied The warranty provided herein and the data specifications and descriptions of this MACHEREY NAGEL product appearing in MACHEREY NAGEL published catalogues and product literature are MACHEREY NAGEL s sole representations concerning the product and warranty No other statements or representations written or oral by MACHEREY NAGEL s employees agent or representatives except written statements signed by a duly authorized officer of MACHEREY NAGEL are authorized they should not be relied upon by the customer and are not a part of the contract of sale or of this warranty Product claims are subject to change Therefore please contact our Technical Service Team for the most up to date information on MACHEREY NAGEL products You may also contact your local distributor for general scientific information Applications mentioned in MACHEREY NAGEL literature are provided for informational purposes only MACHEREY NAGEL does not warrant that all applications have been tested in MACHEREY NAGEL laboratories using MACHEREY NAGEL products MACHEREY NAGEL does not warrant the correctness of any of those applications Last updated 07 2010 Rev 03 Please contact MACHER
12. e 1 2 Equipment and consumables to be supplied by user Product REF Pack of Magnetic separation system 744900 1 e g NucleoMag SEP see section 2 3 Separation plate for magnetic beads separation 740481 4 e g Square well Block 96 well block with 740481 24 24 2 1 mL square wells Lysis tubes for incubation of samples and lysis M l i 740477 4 sets e g Rack of Tubes Strips 1 set consists 740477 24 24 sets of 1 Rack 12 Strips with 8 tubes 1 2 mL wells each and 12 Cap Strips Elution plate for collecting purified nucleic acids 740486 24 24 e g Elution Plate U bottom 96 well 0 3 mL microtiterplate with 300 uL u bottom wells e g Elution Plate Flat bottom 96 well 0 3 mL microtiterplate with 300 uL flat bottom wells For use of kit on KingFisher 96 instrument e g KingFisher 96 Accessory Kit A Square well Blocks Deep well tip combs 744950 Elution Plates for 4 x 96 NucleoMag 96 Tissue preps using KingFisher 96 platform 1 set MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue 2 Product description 2 1 The basic principle The NucleoMag 96 Tissue procedure is based on reversible adsorption of nucleic acids to paramagnetic beads under appropriate buffer conditions Tissue samples cells or bacteria are Iysed with SDS Proteinase K solution Buffer T1 For the adjustment of the binding conditions under which nucleic acids bind to the paramagnetic be
13. e washing and elution steps Alternatively beads can be resuspended in the buffer by pipetting up and down several times For fully automated use on liquid handling workstations a gripper tool is required the plate is transferred to the magnetic separator for separation of the beads and transferred to the shaker module for resuspension of the beads Movable magnetic systems Separators with moving magnetic pins Magnetic pins rods are moved from one side of the well to the other and vice versa Beads follow this movement and are thus pulled through the buffer during the wash and elution steps Separation takes place when the system stops Automated separators Separators with moving magnets Magnetic beads are transferred into suitable plates or tubes Beads are resuspended from the rod covered magnets Following binding washing or elution beads are collected again with the rod covered magnets and transferred to the next plate or tube 2 4 Adjusting the shaker settings When using a plate shaker for the washing and elution steps the speed settings have to be adjusted carefully for each specific separation plate and shaker to prevent cross contamination from well to well Proceed as follows Adjusting shaker speed for binding and wash steps Load 1000 uL for checking the settings for the binding step or 600 uL for checking the settings for the washing steps dyed water to the wells of the separation plate Place the plate on the shak
14. er and start shaking with a moderate speed setting for 30 seconds Turn off the shaker and check the plate surface for small droplets of dyed water MACHEREY NAGEL 05 2014 Rev 03 7 Genomic DNA from tissue Increase speed setting shake for an additional 30 seconds and check the plate surface for droplets again Continue increasing the speed setting until you observe droplets on top of the separation plate Reduce speed setting check again and use this setting for the washing step Adjusting shaker speed for the elution step Load 100 uL dyed water to the wells of the collection plate and proceed as described above 2 5 Handling of beads Distribution of beads A homogeneous distribution of the magnetic beads to the individual wells of the separation plate is essential for a high well to well consistency Therefore before distributing the beads make sure that the beads are completely resuspended Shake the storage bottle well or place it on a vortexer shortly Premixing magnetic beads with the binding buffer allows easier homogenous distribution of the beads to the individual wells of the separation plate During automation a premix step before aspirating the beads binding buffer mixture from the reservoir is recommended to keep the beads resuspended Magnetic separation time Attraction of the magnetic beads to the magnetic pins depends on the magnetic strength of the magnetic pins the selected separation plate
15. hering PE Foil 740676 50 sheets Rack of Tube Strips 740477 4 sets set consists of 1 Rack 12 Tube Strips 740477 24 24 sets with 8 tubes each and 12 Cap Strips Elution Plate U bottom 740486 24 24 Elution Plate Flat bottom 740673 20 KingFisher 96 Accessory Kit A 744950 1 set set consists of Square well Blocks Deep well tip combs Elution Plates for 4 x 96 NucleoMag 96 Tissue preps using KingFisher 96 platform Visit www mn net com for more detailed product information MACHEREY NAGEL 05 2014 Rev 03 21 Genomic DNA from tissue 6 3 Product use restriction warranty NucleoMag 96 Tissue kit components are intended developed designed and sold FOR RESEARCH PURPOSES ONLY except however any other function of the product being expressly described in original MACHEREY NAGEL product leaflets MACHEREY NAGEL products are intended for GENERAL LABORATORY USE ONLY MACHEREY NAGEL products are suited for QUALIFIED PERSONNEL ONLY MACHEREY NAGEL products shall in any event only be used wearing adequate PROTECTIVE CLOTHING For detailed information please refer to the respective Material Safety Data Sheet of the product MACHEREY NAGEL products shall exclusively be used in an ADEQUATE TEST ENVIRONMENT MACHEREY NAGEL does not assume any responsibility for damages due to improper application of our products in other fields of application Application on the human body is STRICTLY FORBIDDEN The respective user is liable for
16. ion buffer For some separators higher elution volumes might be necessary to cover the whole pellet Elution is possible at room temperature Yield can be increased by 15 20 if elution is performed at 55 C MACHEREY NAGEL 05 2014 Rev 03 9 Genomic DNA from tissue 3 Storage conditions and preparation of working solutions Attention Buffers MB2 MB3 and MB4 contain chaotropic salt Wear gloves and goggles Storage conditions All components of the NucleoMag 96 Tissue kit should be stored at room temperature 18 25 C and are stable for up to one year All buffers are delivered ready to use Before starting any NucleoMag 96 Tissue protocol prepare the following Proteinase K Add the indicated volume of Proteinase Buffer PB to dissolve lyophilized Proteinase K Proteinase K solution is stable at 20 C for at least 6 months NucleoMag 96 Tissue 1 x 96 preps 4x 96 preps 24 x 96 preps REF 744300 1 744300 4 744300 24 Proteinase K 75 mg 4x75 mg 24 x 75 mg lyophilized Add 2 6 mL Add 2 6 mL Add 2 6 mL Proteinase Buffer Proteinase Buffer Proteinase Buffer to each vial to each vial 10 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue 4 Safety instructions The following components of the NucleoMag 96 Tissue kits contain hazardous contents Wear gloves and goggles and follow the safety instructions given in this section GHS classification Only harmful features do no
17. ll Be sure to resuspend the NucleoMag B Beads before removing them from the storage bottle Vortex storage bottle briefly until a homogenous suspension has been formed Separate the magnetic beads against the side of the wells by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnets Remove and discard supernatant by pipetting Note Do not disturb the attracted beads while aspirating the supernatant The magnetic pellet is not visible in this step Remove supernatant from the opposite side of the well Wash with MB3 Remove the Square well Block from the NucleoMag SEP magnetic separator Add 600 uL Buffer MB3 to each well and resuspend the beads by shaking until the beads are resuspended completely 5 min Alternatively resuspend beads completely by repeated pipetting up and down 15 times Separate the magnetic beads by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnet Remove and discard supernatant by pipetting Wash with MB4 Remove the Square well Block from the NucleoMag SEP magnetic separator Add 600 pL Buffer MB4 to each well and resuspend the beads by shaking until the beads are resuspended completely 5 min Alternatively resuspend beads completely by repeated pipetting up and down 15 times Separate the magnetic beads
18. n at 55 C for 10 min 18 MACHEREY NAGEL 05 2014 Rev 03 NucleoMag 96 Tissue 6 Appendix 6 1 Troubleshooting Problem Possible cause and suggestions Poor DNA yield Elution buffer volume insufficient Beads pellet must be covered completely with elution buffer Insufficient performance of elution buffer during elution step Remove residual buffers during the separation steps completely Remaining buffers decrease efficiency of following wash steps and elution step Beads dried out Do not let the beads dry as this might result in lower elution efficiencies Partial elution in Wash Buffer MB5 already Keep the beads on the magnet while dispensing Wash Buffer MB5 Do not resuspend beads in this buffer and do not incubate beads in this buffer for more than 2 min as this buffer is water based and might elute the DNA already Aspiration of attracted bead pellet e Do not disturb the attracted beads while aspirating the supernatant especially when the magnetic pellet is not visible in the lysate Incubation after dispensing beads to lysate Mix immediately after dispensing NucleoMag B Beads Buffer MB2 to the lysate Low purity Insufficient washing procedure Use only the appropriate combinations of separator and plate for example Square well Block in combination with NucleoMag SEP Make sure that beads are resuspended completely during the washing procedure If shaking is not s
19. ol is designed for magnetic separators with static pins e g NucleoMag SEP and suitable plate shakers see section 2 3 It is recommended using a Square well Block for separation see section 1 2 Alternatively isolation of DNA can be performed in reaction tubes with suitable magnetic separators This protocol is for manual use and serves as a guideline for adapting the kit to robotic instruments Before starting the preparation Check if Proteinase K was prepared according to section 3 Lyse samples Calculate the amount of lysis stock required for each sample 25 pL of Proteinase K solution 200 pL Buffer T1 are required Prepare lysis stock solution accordingly and vortex Never prepare the lysis stock solution more than 15 min before addition to the samples Proteinase K tends to self digestion when incubated in Buffer T1 without substrate Transfer 225 uL of the resulting stock solution to each lysis tube containing up to 20 mg of tissue sample e g mouse tail section or up to 1 x 10 cultured cells or up to 1 mL of an overnight culture of bacteria Close the individual tubes Mix by vigorous shaking for 10 15 s Spin briefly 15 s 1 500 x g to collect any sample at the bottom of the tube The sample must be submerged in the solution Incubate the tubes containing the samples at 56 C until complete lysis is obtained at least 1 3 h or overnight For cultured cells incubation can be carried out at 70 C for 10 15
20. omic DNA from suitable samples up to 20 mg tissue up to 1x 10 cells or up to 1 mL of an overnight culture of bacteria with an Azgo Azgo ratio 1 6 1 9 and typical concentration of 20 50 ng uL Depending on the elution volume used concentrations of 10 150 ng uL can be obtained Following lysis of samples with Proteinase K NucleoMag 96 Tissue can be processed completely at room temperature however elution at 55 C will increase the yield by about 15 20 NucleoMag B Beads are highly reactive superparamagnetic beads The binding capacity is 0 4 ug of gDNA per 1 uL of NucleoMag B Bead Suspension 1 uL of suspension contains 130 ug of beads 6 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue 2 3 Magnetic separation systems For use of NucleoMag 96 Tissue the use of the magnetic separator NucleoMag SEP is recommended Separation is carried out in a Square well Block see ordering information The kit can also be used with other common separators Magnetic separator Separation plate or tube NucleoMag SEP MN REF 744900 Square well Block MN REF 740481 24 Tecan Te MagS 1 5 mL tubes without lid Sarstedt Static magnetic pins Separators with static magnetic pins for example NucleoMag SEP for manual use and for use on liquid handling workstations This type of separator is recommended in combination with a suitable microplate shaker for optimal resuspension of the beads during th
21. t need to be labeled with H and P phrases up to 125 mL or 125 g Mindergef hrliche Eigenschaften m ssen bis 125 mL oder 125 g nicht mit H und P S tzen gekennzeichnet werden Component Hazard contents GHS symbol Hazard Precaution phrases phrases Inhalt Gefahrstoff GHS Symbol H S tze P S tze MB2 Sodium perchlorate 20 Warning 226 302 210 233 301 312 40 ethanol 35 55 D 330 403 235 Natriumperchlorat 20 40 Achtung Ethanol 35 55 I MB3 MB4 Sodium perchlorate amp Warning 226 210 233 403 235 5 20 ethanol 20 35 Natriumperchlorat 5 20 Achtung Ethanol 20 35 Proteinase K Proteinase K Iyophilized Proteinase K Iyophilisiert Danger 315 319 261 280 302 352 Gefahr 334 335 304 340 305 351 338 312 333 313 337 313 342 311 403 233 OO Hazard phrases H 226 Flammable liquid and vapour Fl ssigkeit und Dampf entz ndbar H 302 Harmful if swallowed Gesundheitssch dlich bei Verschlucken H 315 Causes skin irritation Verursacht Hautreizungen H 319 Causes serious eye irritation Verursacht schwere Augenreizung H 334 May cause allergy or asthma symptoms or breathing difficulties if inhaled Kann bei Einatmen Allergie asthmaartige Symptome oder Atembeschwerden verursachen H 335 May cause respiratory irritation Kann die Atemwege reizen MACHEREY NAGEL 05 2014 Rev 03 11 Genomic DNA from tissue Precaution phrases P 210 P 233 P 2
22. tions MACHEREY NAGEL s sole obligation and the customer s sole remedy is limited to replacement of products free of charge in the event products fail to perform as warranted Supplementary reference is made to the general business terms and conditions of MACHEREY NAGEL which are printed on the price list Please contact us if you wish to get an extra copy There is no warranty for and MACHEREY NAGEL is not liable for damages or defects arising in shipping and handling transport insurance for customers excluded or out of accident or improper or abnormal use of this product defects in products or 22 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue components not manufactured by MACHEREY NAGEL or damages resulting from such non MACHEREY NAGEL components or products MACHEREY NAGEL makes no other warranty of any kind whatsoever and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WARRANTIES OF ANY KIND OR NATURE WHATSOEVER DIRECTLY OR INDIRECTLY EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION AS TO THE SUITABILITY REPRODUCTIVITY DURABILITY FITNESS FOR A PARTICULAR PURPOSE OR USE MERCHANTABILITY CONDITION OR ANY OTHER MATTER WITH RESPECT TO MACHEREY NAGEL PRODUCTS In no event shall MACHEREY NAGEL be liable for claims for any other damages whether direct indirect incidental compensatory foreseeable consequential or special including but not limited to loss of use revenue or profit whether based upon warr
23. ufficient to resuspend the beads completely mix by repeated pipetting up and down MACHEREY NAGEL 05 2014 Rev 03 19 Genomic DNA from tissue Problem Possible cause and suggestions Carry over of ethanol wash solutions Suboptimal 3 i Be sure to remove all of the ethanolic wash solution as per ormance residual ethanol interferes with downstream applications of DNA in downstream ahead Low purity applications See above Carry over of beads Time for magnetic separation too short Increase separation time to allow the beads to be completely attracted to the magnetic pins before aspirating any liquid from the well Aspiration speed too high elution step High aspiration speed during the elution step may cause bead carry over Reduce aspiration speed for elution step Cross contamination Contamination of the rims Do not moisten the rims of the Square well Block when transferring the tissue lysate If the rim of the wells is contaminated seal the Square well Block with Self adhering PE Foil see ordering information before starting the shaker 20 MACHEREY NAGEL 05 2014 Rev 03 Genomic DNA from tissue 6 2 Ordering information Product REF Pack of NucleoMag 96 Tissue 744300 1 1 x 96 preps 744300 4 4 x 96 preps 744300 24 24 x 96 preps Buffer T1 740940 25 25 mL RNase A 740505 50 50 mg NucleoMag SEP 744900 1 Square well Blocks 740481 4 740481 24 24 Self ad
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