Qiagen TissueLyser PDF
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1. E Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions E f using a QIAGEN kit for DNA purification read the supplied handbook and appropriate supplementary protocol carefully before starting Procedure l Place the tissues in 2 ml microcentrifuge tubes containing 1 stainless steel bead 5 mm mean diameter 2 Add the appropriate volume of lysis buffer e g Buffer ATL to each tube 3 Place the tubes in the TissueLyser Adapter Set 2 x 24 4 Operate the TissueLyser for 20 s at 15 Hz Note Exceeding this homogenization time and intensity may lead to significant fragmentation of genomic DNA If working with fibrous tissues cutting the tissue into smaller pieces before starting disruption will improve disruption efficiency 5 Proceed with DNA purification 22 Tissuelyser Handbook 10 2010 Protocol Purification of DNA from Plant Tissues Mini Protocol This protocol provides guidelines on using Tissuelyser Adapter Sets to disrupt plant tissues for subsequent DNA purification If using a GIAGEN kit for DNA purification see Table 5 page 10 refer to the supplied handbook which contains a complete protocol for sample disruption and DNA purification Important points before starting MH Before beginning the procedure read Important Notes page 13 E Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions BW Ifusing a QIAGEN kit2. and disruption under frozen conditions Procedure 1 If handling frozen tissues precool the TissueLyser Adapter Set 2 x 24 or TissueLyser Adapter Set 2 x 96 by storing at 80 C for at least 2 h The adapter sets do not need to be precooled if handling fresh tissues 2 Place the tissues in 2 ml microcentrifuge tubes or 1 2 ml collection microtubes containing 1 stainless steel bead 3 7 mm mean diameter If handling frozen tissues keep the tubes on dry ice Note Do not freeze the tubes in liquid nitrogen as this may lead to breakage of the tubes 3 Immediately add the appropriate volume of lysis buffer e g Buffer RLT or Buffer RLC to each tube If handling frozen tissues do not add lysis buffer Note Do not use Buffer RLT or Buffer RLC with tungsten carbide beads as these buffers can react with and damage the bead surface 4 Place the tubes in the TissueLyser Adapter Set 2 x 24 if using 2 ml tubes or the Tissuelyser Adapter Set 2 x 96 if using 1 2 ml tubes If using the RNeasy 96 Kit refer to supplementary protocol Isolation of total RNA from plants using the RNeasy 96 Kit RY23 18 Tissuelyser Handbook 10 2010 5 Operate the TissueLyser for 1 min at 30 Hz Disassemble the adapter set rotate the rack of tubes so that the tubes nearest to the TissueLyser are now outermost and reassemble the adapter set Operate the TissueLyser for another 1 min at 30 Hz The duration of disruption and homogenization depen
3. columns When disrupting and homogenizing tissues in Buffer RLT Plus excessive foaming may occur This foaming is substantially reduced by adding Reagent DX to Buffer RLT Plus at a final concentration of 0 5 v v before starting disruption and homogenization Reagent DX has been carefully tested with RNeasy Plus Kits and AllPrep Kits and has no effect on RNA purity or on downstream applications such as realtime RT PCR Buffer RLT Plus containing Reagent DX can be stored at room temperature 15 25 C for at least 9 months Reagent DX is supplied separately for ordering information see page 32 Tissuelyser Handbook 10 2010 15 ES ES I zt o EZ E32 at LS Protocol Purification of RNA or Multiple Analytes from Animal and Human Tissues This protocol provides guidelines on disrupting animal and human tissues for purification of RNA or for simultaneous purification of DNA and RNA or DNA RNA and protein If using a QIAGEN sample purification kit see Tables 1 2 and 6 on pages 7 8 and 10 refer to the supplied handbook which contains a complete protocol for sample disruption and purification Important points before starting Before beginning the procedure read Important Notes page 13 Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions If using a QIAGEN sample purification kit read the supplied handbook carefully before starting After storage in RNAlater RNA Sta
4. continue using DNeasy Kits QlAamp Kits RNeasy Kits and the miRNeasy Mini Kit for purification of high quality DNA or RNA For more information about the automated procedure see the relevant protocol sheet available at www giagen com MyQlAcube The QlAcube The QlAcube is preinstalled with protocols for purification of plasmid DNA genomic DNA RNA viral nucleic acids and proteins plus DNA and RNA cleanup The range of protocols available is continually expanding and additional QIAGEN protocols can be downloaded free of charge at www giagen com MyQ lAcube Automated purification using magnetic particles and 96 well plates Complete automated solutions from QIAGEN allow purification of genomic DNA or total RNA from human animal or plant tissues at a range of different throughputs using magnetic particles or 96 well plates see Table 7 page 30 QIAGEN Instrument Service provides comprehensive support services to ensure the continued success of your automated applications For more information about QIAGEN automation and QIAGEN Instrument Service visit www qiagen com automation Tissuelyser Handbook 10 2010 29 Table 7 Automated purification of genomic DNA and total RNA from tissues Workstation Capability EZ1 Advanced Purification of genomic DNA or total RNA from 1 6 human samples per run QlAsymphony SP Purification of genomic DNA or total RNA from 1 96 animal or human samples per run BioRobot Universal Purifica
5. easy to lyse tissues on the BioRobot M48 workstation MagAttract RNA Tissue For 192 preps MagAttract 959236 Mini M48 Kit 192 Suspension E DNase I Buffers MagAttract RNA Universal Tissue M48 Kit for purification of total RNA from all types of tissue on the BioRobot M48 workstation MagAttract RNA Universal For 192 preps MagAttract 956336 Tissue M48 Kit 192 Suspension E DNase QIAzol Lysis Reagent Buffers GlAsymphony RNA Kit for purification of total RNA from cells and tissues on the GlAsymphony SP QlAsymphony RNA Kit 192 For 192 preps 2 Reagent 931636 Cartridges and Enzyme Racks Tissuelyser Handbook 10 2010 33 Ordering Information Product Contents Cat no RNeasy Plant Mini Kit for purification of total RNA from plants and fungi RNeasy Plant Mini Kit 20 For 20 preps RNeasy Spin Columns 74903 QlAshredder Spin Columns Collection Tubes Buffers RNeasy 96 Kit for purification of total RNA from cells in 96 well format RNeasy 96 Kit 4 For 4 x 96 preps RNeasy 96 Plates 74181 Elution Microtubes CL Caps S Blocks Airpore Tape Sheets Buffers RNeasy Protect Bacteria Kits for stabilization and purification of total RNA from bacteria RNeasy Protect Bacteria For 50 preps RNAprotect Bacteria 74524 Mini Kit 50 Reagent RNeasy Mini Kit RNeasy Protect Bacteria For 10 preps RNAprotect Bacteria 72555 Midi Kit 10 Reagent RNeasy Midi Kit AllPrep DNA RNA Protein Mini Kit for simultaneous purification
6. for DNA purification read the supplied handbook carefully before starting Bb Fresh frozen or lyophilized tissues can be processed Fresh tissues can be disrupted in lysis buffer at ambient temperature Alternatively fresh or frozen tissues can be disrupted without lysis buffer if they are precooled on dry ice and if the adapter sets are precooled at 80 C for at least 2 h Lyophilized tissues can be disrupted without lysis buffer at ambient temperature Disruption of tissues in lysis buffer yields DNA ideal for PCR while disruption of tissues in liquid nitrogen yields DNA of a higher molecular weight We do not recommend disrupting frozen tissues in lysis buffer as this results in low yields and degraded DNA Procedure l lf purifying DNA of higher molecular weight from fresh or frozen tissues precool the TissueLyser Adapter Set 2 x 24 or TissueLyser Adapter Set 2 x 96 by storing at 80 C for at least 2 h The adapter sets do not need to be precooled if disrupting fresh tissues in lysis buffer or if disrupting lyophilized tissues 2 Place the tissues in 2 ml microcentrifuge tubes or 1 2 ml collection microtubes containing 1 tungsten carbide bead 3 mm mean diameter 3 f purifying DNA of higher molecular weight from fresh or frozen tissues precool the tubes by storing on dry ice Note Do not freeze the tubes in liquid nitrogen as this may lead to breakage of the tubes The tubes do not need to be precooled if disrupting fresh tissue
7. run Also automatable on BioRobot EZ1 Tissuelyser Handbook 10 2010 9 Table 5 Kits for DNA purification from plant tissues Kit Kit format Page DNeasy Plant Spin column up to 100 mg tissue automatable on 23 Mini Kit GlAcube DNeasy Plant Spin column up to 1 g tissue 29 Maxi Kit DNeasy 96 96 well plate up to 50 mg tissue 23 Plant Kit MagAttract 96 Magnetic particles up to 100 mg tissue automatable 2S DNA Plant on BioRobot Plant Science System Genotyping Core Kit BioSprint 15 Magnetic particles up to 50 mg tissue automated on 23 DNA Plant Kit BioSprint 15 up to 15 samples per run BioSprint 96 Magnetic particles up to 50 mg tissue automated 23 DNA Plant Kit on BioSprint 96 up to 96 samples per run No longer available Table 6 Kits for simultaneous purification of multiple analytes from animal or human tissues Analytes purified Kit Kit format Page DNA RNA and AllPrep DNA RNA Spin column up to 30 mg 16 protein Protein Mini Kit tissue DNA and RNA AllPrep DNA RNA Spin column up to 5 mg 16 Micro Kit tissue AllPrep DNA RNA Spin column up to 30 mg 16 Mini Kit tissue 10 Tissuelyser Handbook 10 2010 GIAGEN Supplementary Protocols Many of the protocols listed in this handbook are supplementary to the protocols found in the handbook of the specific kit being used QIAGEN is constantly developing new protocols for existing products These supplementary protocols ca
8. 31235 Mini Kit 96 2 Reagent Cartridges and Enzyme Racks GlAsymphony DNA For 96 preps of 1000 pl each QoS Midi Kit 96 2 Reagent Cartridges and Enzyme Racks DNeasy Blood amp Tissue Kit for purification of total DNA from animal blood and tissues and from cells yeast bacteria or viruses DNeasy Blood amp Tissue Kit 50 For 50 preps DNeasy Spin Columns 69504 Collection Tubes Proteinase K Buffers DNeasy 96 Blood amp Tissue Kit for purification of total DNA from animal blood and tissues and from cells yeast bacteria or viruses in 96 well format DNeasy 96 Blood amp For 4 x 96 preps DNeasy 96 Plates 69581 Tissue Kit 4 Collection Microtubes Caps S Blocks Elution Microtubes RS AirPore Tape Sheets Proteinase K Buffers DNeasy Plant Kits for purification of total DNA from plants and fungi DNeasy Plant Mini Kit 50 For 50 preps DNeasy Spin Columns 69104 QlAshredder Spin Columns Collection Tubes RNase A Buffers Tissuelyser Handbook 10 2010 35 Ordering Information Product Contents Cat no DNeasy Plant Maxi Kit 6 For 6 preps DNeasy Spin Columns 68161 QlAshredder Spin Columns Collection Tubes RNase A Buffers DNeasy 96 Plant Kit for purification of total DNA from plants in 96 well format DNeasy 96 Plant Kit 6 For 6 x 96 preps DNeasy 96 Plates 69181 Collection Microtubes Caps S Blocks Elution Microtubes RS AirPore Tape Sheets RNase A Reagent DX Buffers MagAttract 96 DNA Plant Co
9. Homogenization of the material acts to shear the high molecular weight cellular proteins and carbohydrates that may otherwise reduce binding of DNA and RNA to silica membranes or magnetic particles Sample disruption using for example a mortar and pestle does not result in efficient homogenization The TissueLyser both disrupts and homogenizes sample material in one simple and reliable step The Tissuelyser is easily programmed to provide variable speeds from 3 to 30 Hz 180 1800 oscillations minute and run times from 10 seconds to 99 minutes Applications The ability to process up to 192 samples per run makes the Tissuelyser the ideal front end solution to access biological information for genomics transcriptomics and proteomics applications For nextgeneration high throughput sequencing technologies such as polony sequencing the Tissuelyser is the disruption instrument of choice The Tissuelyser enables fast and uniform disruption of animal and human tissues plant tissues bacteria and yeast in various sample volumes in several formats QIAGEN offers adapter sets for 2 x 96 collection microtubes 1 2 ml or 2 x 24 microcentrifuge tubes 2 ml as well as stainless steel and tungsten carbide beads For disruption of large samples grinding jar sets 10 ml with stainless steel or Teflon grinding balls are also available from QIAGEN For more details about these and other accessories for the Tissuelyser see Appendix A page 27 The Tiss
10. Second Edition October 2010 TissueLyser Handbook For high throughput disruption of biological samples GIAGEN Sample and Assay Technologies GIAGEN is the leading provider of innovative sample and assay technologies enabling the isolation and detection of contents of any biological sample Our advanced high quality products and services ensure success from sample to result QIAGEN sets standards in Purification of DNA RNA and proteins Nucleic acid and protein assays microRNA research and RNAi Automation of sample and assay technologies Our mission is to enable you to achieve outstanding success and breakthroughs For more information visit www giagen com Contents Product Contents 4 Storage 4 Product Use Limitations 4 Product Warranty and Satisfaction Guarantee 4 Technical Assistance 5 Safety Information 5 Introduction 6 Principle 6 Applications 6 QIAGEN Supplementary Protocols 11 Equipment and Reagents to Be Supplied by User 12 Important Notes 13 General remarks on disruption and homogenization 13 Disruption and homogenization using the Tissuelyser 13 Disruption and homogenization in Buffer RLT Plus 15 Protocols Purification of RNA or Multiple Analytes from Animal and Human Tissues 16 B6 Purification of RNA from Plant Tissues 18 E Purification of RNA from Bacteria 20 Purification of RNA from Yeast 21 Purification of DNA from Animal and Human Tissues 22 Purification of DNA from Plant Tissues Mini Pro
11. an misuse QIAGEN will replace it free of charge or refund the purchase price We reserve the right to change alter or modify any product to enhance its performance and design If a QIAGEN product does not meet your expectations simply call your local Technical Service Department or distributor We will credit your account or exchange the product as you wish Separate conditions apply to QIAGEN scientific instruments service products and to products shipped on dry ice Please inquire for more information 4 Tissuelyser Handbook 10 2010 A copy of QIAGEN terms and conditions can be obtained on request and is also provided on the back of our invoices If you have questions about product specifications or performance please call QIAGEN Technical Services or your local distributor see back cover or visit www qiagen com Technical Assistance At QIAGEN we pride ourselves on the quality and availability of our technical support Our Technical Service Departments are staffed by experienced scientists with extensive practical and theoretical expertise in sample and assay technologies and the use of QIAGEN products If you have any questions or experience any difficulties regarding the TissueLyser or QIAGEN products in general please do not hesitate to contact us QIAGEN customers are a major source of information regarding advanced or specialized uses of our products This information is helpful to other scientists as well as to the rese
12. archers at QIAGEN We therefore encourage you to contact us if you have any suggestions about product performance or new applications and techniques For technical assistance and more information please see our Technical Support Center t www giagen com Support or call one of the QIAGEN Technical Service Departments or local distributors see back cover or visit www giagen com Safety Information When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information please consult the appropriate material safety data sheets MSDSs These are available online in convenient and compact PDF format at www qiagen com Support MSDS aspx where you can find view and print the MSDS for each QIAGEN kit and kit component 24 hour emergency information Emergency medical information in English French and German can be obtained 24 hours a day from Poison Information Center Mainz Germany Tel 49 6131 19240 Tissuelyser Handbook 10 2010 5 Introduction Principle The TissueLyser provides rapid and efficient disruption of up to 192 biological samples including animal and human tissues plant tissues bacteria and yeast Disruption and homogenization are achieved through the beating and grinding effect of beads on the sample material as they are shaken together in the grinding vessels Disruption is critically important in order to release the nucleic acids from the sample material
13. atents GlAzol Lysis Reagent is a subject of US Patent No 5 346 994 and foreign equivalents 2004 2010 QIAGEN all rights reserved www qiagen com Australia Orders 1 800 243 800 Fax 03 9840 9888 Technical 1 800 243 066 Austria Orders 0800 28 10 10 Fax 0800 28 10 19 Technical 0800 28 10 11 Belgium Orders 0800 79612 Fax 0800 79611 Technical 0800 79556 Brazil Orders 0800 557779 Fax 55 11 5079 4001 Technical 0800 557779 Canada Orders 800 572 9613 Fax 800 713 5951 Technical 800 DNA PREP 800 362 7737 China Orders 86 21 3865 3865 Fax 86 21 3865 3965 Technical 800 988 0325 Denmark Orders 80 885945 Fax 80 885944 Technical 80 885942 Finland Orders 0800 914416 Fax 0800 914415 Technical 0800 914413 France Orders 01 60 920 926 Fax 01 60 920 925 Technical 01 60 920 930 Offers 01 60 920 928 Germany Orders 02103 29 12000 Fax 02103 29 22000 Technical 02103 29 12400 Hong Kong Orders 800 933 965 Fax 800 930 439 Technical 800 930 425 Ireland Orders 1800 555 049 Fax 1800 555 048 Technical 1800 555 061 Italy Orders 800 789 544 Fax 02 334304 826 Technical 800 787980 Japan Telephone 03 6890 7300 Fax 03 5547 0818 Technical 03 6890 7300 Korea South Orders 080 000 7146 Fax 02 2626 5703 Technical 080 000 7145 Luxembourg Orders 8002 2076 Fax 8002 2073 Technical 8002 2067 Mexico Orders 01 800 7742 639 Fax 01 800 1 122 330 Technical 01 800 7742 436 The Netherla
14. bilization Reagent or Allprotect Tissue Reagent tissues become slightly hard If disrupting in Buffer RLT we recommend increasing the volume of this buffer according to the protocols in the RNeasy Mini Handbook In addition the disruption time may be need to be extended Procedure 1 Place the tissues in 2 ml microcentrifuge tubes or 1 2 ml collection microtubes containing 1 stainless steel bead 3 7 mm mean diameter If handling fresh or frozen tissue samples keep the tubes on dry ice Place the tubes at room temperature 15 25 C Immediately add the appropriate volume of lysis buffer e g Buffer RLT Buffer RLT Plus or QIAzol Lysis Reagent to each tube Note Do not use Buffer RLT Buffer RLT Plus or QIAzol Lysis Reagent with tungsten carbide beads as these buffers can react with and damage the bead surface Note If using Buffer RLT Plus we recommend adding Reagent DX to prevent excessive foaming For details see Disruption and homogenization in Buffer RLT Plus page 15 Place the tubes in the TissueLyser Adapter Set 2 x 24 if using 2 ml tubes or the TissueLyser Adapter Set 2 x 96 if using 1 2 ml tubes Tissuelyser Handbook 10 2010 4 Operate the TissueLyser for 2 min at 20 30 Hz Disassemble the adapter set rotate the rack of tubes so that the tubes nearest to the TissueLyser are now outermost and reassemble the adapter set Operate the TissueLyser for another 2 min at 20 30 Hz The duration of disrupti
15. cts and qualitative and quantitative RNA analysis is now easier and faster than ever To find out more visit www giagen com QlAxcel 30 Tissuelyser Handbook 10 2010 Ordering Information Product Contents Cat no TissueLyser Il Universal laboratory mixer mill 85300 disruptor 100 120 220 240 V 50 60 Hz Accessories Tissuelyser Adapter Set 2 x 24 2 sets of Adapter Plates and 2 racks 69982 for use with 2 ml microcentrifuge tubes on the TissueLyser Tissuelyser Adapter Set 2 x 96 2 sets of Adapter Plates for use with 69984 Collection Microtubes racked on the TissueLyser Grinding Jar Set 2 Grinding Jars 10 ml 2 Stainless 69985 S Steel 2 x 10 ml Steel Grinding Balls 20 mm Grinding Jar Set 2 Grinding Jars 10 ml 2 Teflon 69986 Teflon 2 x 10 ml Grinding Balls 20 mm Stainless Steel Beads Stainless Steel Beads suitable for 69989 5 mm 200 use with the TissueLyser system Tungsten Carbide Beads Tungsten Carbide Beads suitable for 69997 3 mm 200 use with the TissueLyser system TissueLyser Single Bead For dispensing individual beads 69965 Dispenser 5 mm 5 mm diameter TissueLyser Single Bead For dispensing individual beads 69967 Dispenser 7 mm 7 mm diameter Tissuelyser 3 mm Bead For dispensing 96 beads 3 mm 69973 Dispenser 96 Well diameter in parallel Tissuelyser 5 mm Bead For dispensing 96 beads 5 mm 69975 Dispenser 96 Well diameter in parallel Collection Microtubes racked Nonsterile poly
16. ds on the tissue being processed and can be extended until no tissue debris is visible If necessary keep the samples on dry ice for several minutes in between the individual disruption steps to avoid thawing of the samples Rearranging the tubes ensures uniform disruption and homogenization 6 Proceed with RNA purification If frozen samples were disrupted add lysis buffer and proceed with RNA purification E Z z s nss j 4UD d Do not reuse the stainless steel beads Tissuelyser Handbook 10 2010 19 Ed z E g E Y io en Protocol Purification of RNA from Bacteria This protocol provides guidelines on disrupting bacteria for subsequent RNA purification If using an RNeasy Protect Bactera Kit for RNA purification see Table 3 page 9 refer to the supplied RNAprotect Bacteria Reagent Handbook which contains complete protocols for sample disruption and RNA purification Important points before starting E Before beginning the procedure read Important Notes page 13 E Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions E f using an RNeasy Protect Bacteria Kit for RNA purification read the supplied handbook carefully before starting E Bead milling will disrupt most Gram positive and Gram negative bacteria including mycobacteria Gram positive bacteria usually require more rigorous digestion e g increased enzyme digestion time and temperature a
17. e of lysis buffer e g Buffer RLT to each sample and vortex vigorously 2 Transfer each sample to 2 ml microcentrifuge tubes containing 600 pl acid washed glass beads 450 550 pm mean diameter 3 Place the tubes in the TissueLyser Adapter Set 2 x 24 4 Operate the TissueLyser for 5 min at 30 Hz The duration of disruption and homogenization depends on the sample being processed and can be extended until no debris is visible 5 Proceed with RNA purification Tissuelyser Handbook 10 2010 21 VNA 15094 Protocol Purification of DNA from Animal and Human Tissues This protocol provides guidelines on disrupting animal and human tissues for subsequent DNA purification If using a QIAGEN kit for DNA purification see Table 4 page 9 refer to the following supplementary protocols for the complete procedure for sample disruption and DNA purification DNeasy Blood amp Tissue Kit Purification of total DNA from soft tissues using the Tissuelyser and the DNeasy Blood amp Tissue Kit DY11 QlAamp DNA Mini Kit Isolation of DNA from soft tissues using the Tissuelyser and QlAamp DNA Mini Kit QA31 EZ1 DNA Tissue Kit Isolation of DNA from soft tissue using the Tissuelyser and EZ1 DNA Tissue Kit MA23 MagAttract DNA Mini M48 Kit Isolation of DNA from soft tissue using the Tissuelyser and MagAttract Mini M48 Kit MA22 Important points before starting E Before beginning the procedure read Important Notes page 13
18. fore significantly reduced DNA and RNA yields Cellular disruption is one of the most critical steps in nucleic acid purification Disruption in lysis buffer alone without physical shearing may result in nucleic acid degradation by endogenous DNases and RNases Incomplete disruption prevents the lysis buffer which inactivates nucleases from contacting nucleic acids within the intact cells Furthermore cellular debris that is not disrupted can result in decreased yield and increases the risk of clogging the purification column After sample disruption there should be no visible particulates except when disrupting materials containing hard noncellular components such as connective tissue bone or woody plant tissue QIAGEN kits and protocols contain recommendations for the most appropriate method of sample disruption and homogenization to maximize the yield and quality of your DNA and RNA preparation Disruption and homogenization using the TissueLyser In bead milling cells and tissues can be disrupted by rapid agitation in the presence of beads Disruption and simultaneous homogenization occur by the shearing and crushing action of the beads as they collide with the sample Disruption efficiency is influenced by E Size and composition of beads Ratio of buffer to samples if buffer is used Amount of starting material Configuration of Tissuelyser i e speed and duration Consistency of sample Type of disruption vessel T
19. g frozen grinding jar sets or if disrupting lyophilized tissues 4 Operate the Tissuelyser for 1 min at 30 Hz 5 If purifying DNA of higher molecular weight from fresh or frozen tissues freeze the jars in liquid nitrogen for 1 min The grinding jars do not need to be frozen if disrupting fresh tissues in lysis buffer or if disrupting lyophilized tissues Tissuelyser Handbook 10 2010 25 o x 3 A gt o JI 6 Operate the TissueLyser for 1 min at 30 Hz 7 Add lysis buffer e g Buffer AP1 if necessary and proceed with DNA purification The stainless steel grinding balls can be reused For details on recovering and cleaning grinding balls refer to the DNeasy Plant Handbook Sz 2z 20 m x 3 a 26 Tissuelyser Handbook 10 2010 Appendix A TissueLyser Accessories Tissuelyser Adapter Set 2 x 24 This adapter set allows disruption of 48 2 x 24 samples in parallel using standard 2 ml microcentrifuge tubes e g Eppendorf Safe lock micro test tubes Sample disruption can be carried out at room temperature or after storing the adapter set at 80 C for at least 2 hours The adapter set can be cleaned with detergent microbicides or up to 96 ethanol For more information see the product sheet sup plied with the Tissuelyser Adapter Set TissueLyser Adapter Set 2 x 96 This adapter set allows disruption of 192 2 x 96 samples in parallel using Collection Microtubes racked Sample d
20. ion Microtube Caps El Optional Tissuelyser 3 mm Bead Dispenser 96 Well or TissueLyser 5 mm Bead Dispenser 96 Well Disruption of 2 large samples MH For disruption of hard samples and disruption in liquid nitrogen Grinding Jar Set S Steel MH For disruption of most samples Grinding Jar Set Teflon See page 31 for ordering information t This is not a complete list of suppliers and does not include many important vendors of biological supplies P pp y Imp g Pp 12 Tissuelyser Handbook 10 2010 Important Notes General remarks on disruption and homogenization Efficient disruption and homogenization of the starting material is an absolute requirement for all nucleic acid purification procedures Disruption and homogenization are 2 distinct steps Disruption Complete disruption of cell walls and plasma membranes of cells and organelles is absolutely required to release all the nucleic acids contained in the sample Different samples require different methods to achieve complete disruption Incomplete disruption results in significantly reduced DNA and RNA yields BH Homogenization Homogenization is necessary to reduce the viscosity of the cell lysates produced by disruption Homogenization shears the high molecular weight cellular proteins and carbohydrates to create a homogeneous lysate Incomplete homogenization results in inefficient binding of nucleic acids to QIAGEN silica membranes and magnetic particles and there
21. isruption can be carried out at room temperature or after storing the adapter set at 80 C for at least 2 hours The adapter set can be cleaned with detergent microbicides or up to 9676 ethanol For more information see the product sheet supplied with the Tissuelyser Adapter Set Tissuelyser Single Bead Dispenser 5 mm This bead dispenser dispenses individual beads 5 mm diameter into any sample container The reservoir holds approximately 150 beads The Tissuelyser Single Bead Dispenser can be cleaned with water or detergent For more information see the product sheet supplied with the Tissuelyser Single Bead Dispenser TissueLyser Single Bead Dispenser 7 mm This bead dispenser dispenses individual beads 7 mm diameter into any sample container The reservoir holds approximately 45 beads The Tissuelyser Single Bead Dispenser can be cleaned with water or detergent For more information see the product sheet supplied with the Tissuelyser Single Bead Dispenser Tissuelyser 3 mm Bead Dispenser 96 well This bead dispenser dispenses 96 beads 3 mm diameter in parallel into Collection Microtubes racked enabling high throughput disruption and homogenization The reservoir holds approximately 1000 beads The dispenser can be cleaned with water or detergent For more information see the product sheet supplied with the Tissuelyser Bead Dispenser 96 well Tissuelyser Handbook 10 2010 27 TissueLyser 5 mm Bead Dispenser 96 well Thi
22. issuelyser Handbook 10 2010 13 Disruption and homogenization methods When using the Tissuelyser in combination with QIAGEN sample purification kits one of 2 methods for disruption and homogenization is carried out samples are either disrupted and homogenized in lysis buffer at room temperature or precooled and then disrupted and homogenized without lysis buffer With the latter method lysis buffer is added after disruption and homogenization The method of precooling samples depends on the Tissuelyser accessory used If using the Tissuelyser Adapter Set 2 x 24 or Tissuelyser Adapter Set 2 x 96 the adapter set should be stored at 80 C for at least 2 hours prior to starting disruption and homogenization and the tubes containing the samples should be precooled on dry ice If using a Grinding Jar Set the jar containing the sample can be frozen in liquid nitrogen prior to starting disruption and homogenization Important When using a Tissuelyser Adapter Set do not freeze the adapter set or the sample tubes in liquid nitrogen as this may result in breakage of the tubes In special cases e g the disruption of teeth or plant seeds the sample can be disrupted and homogenized at room temperature without lysis buffer although this increases the risk of nucleic acid degradation by nucleases Bead selection For disruption of small samples the optimal beads to use are 0 1 0 6 mm mean diameter glass beads for bacteria 0 5 mm glass bead
23. iversal System up to 80 mg tissue miRNeasy 96 Kit 96 well plate up to 100 mg tissue 16 Also automatable on BioRobot EZ1 Also automatable on BioRobot Gene Expression Real Time RT PCR and BioRobot 8000 8 Tissuelyser Handbook 10 2010 Table 3 Kits for RNA purification from plant tissues bacteria and yeast Sample type Kit Kit format Page Plant tissue RNeasy Plant Spin column up to 100 mg tissue 18 e g leaf Mini Kit automatable on QlAcube RNeasy 96 Kit 96 well plate up to 25 mg tissue 18 Bacteria RNeasy Protect Spin column up to 2 5 x 10 cells 20 Gram Bacteria Mini Kit pasive and RNeasy Protect Spin column up to 1 5 x 10 cells 20 negative Bacteria Midi Kit Yeast RNeasy Mini Kit Spin column up to 5 x 10 cells 21 Table 4 Kits for DNA purification from animal or human tissues Kit Kit format Page DNeasy Blood Spin column up to 25 mg tissue automatable 22 amp Tissue Kit on GlAcube DNeasy 96 96 well plate up to 20 mg tissue 22 Blood amp Tissue Kit QlAamp DNA Spin column up to 25 mg tissue automatable 22 Mini Kit on GlAcube EZ1 DNA Tissue Magnetic particles up to 40 mg tissue automated Do Kit on EZ Advanced 1 6 samples per run MagAttract DNA Magnetic particles up to 40 mg tissue automated 22 Mini M48 Kit on BioRobot M48 6 48 samples per run GlAsymphony Magnetic particles up to 50 mg tissue automated 22 DNA Mini Kit on GlAsymphony SP 1 96 samples per
24. n be obtained by contacting one of the QIAGEN Technical Service Departments or local distributors see back cover or visit www qiagen com or visiting our Technical Support Center at www giagen com Support Supplementary protocols can be identified by their reference number which is made up of 2 letters followed by 2 numbers e g RY23 Isolation of total RNA from plants using the RNeasy 96 Kit Note All protocols for use with the Mixer Mill can be used on the Tissuelyser without modification Tissuelyser Handbook 10 2010 11 Equipment and Reagents to Be Supplied by User When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate material safety data sheets MSDSs available from the product supplier For all protocols Kit for purification of DNA and or RNA see ordering information on pages 31 36 or visit www giagen com E Optional Reagent DX see page 15 for details E Optional Liquid nitrogen or dry ice see the individual protocols Disruption of 2 x 48 samples E Tissuelyser Adapter Set 2 x 24 B 2 ml microcentrifuge tubes e g Eppendorf Safe Lock micro test tubes MH Stainless steel or tungsten carbide beads Oo Optional Tissuelyser Single Bead Dispenser 5 mm or Tissuelyser Single Bead Dispenser 7 mm Disruption of 2 x 96 samples E Tissuelyser Adapter Set 2 x 96 MH Collection Microtubes racked BW Collect
25. nd mechanical treatment than Gram negative bacteria For details see the RNAprotect Bacteria Reagent Handbook Procedure 1 Pellet the bacterial cells by centrifugation Immediately add the appropriate volume of lysis buffer e g Buffer RLT to each sample and vortex vigorously 2 Transfer each sample to 2 ml microcentrifuge tubes containing 25 50 mg acid washed glass beads 150 600 pm mean diameter 3 Place the tubes in the TissueLyser Adapter Set 2 x 24 Operate the TissueLyser for 5 min at 30 Hz The duration of disruption and homogenization depends on the sample being processed and can be extended until no debris is visible 5 Proceed with RNA purification 20 Tissuelyser Handbook 10 2010 Protocol Purification of RNA from Yeast This protocol provides guidelines on disrupting yeast cells for subsequent RNA purification If using the RNeasy Mini Kit for RNA purification see Table 3 page 9 refer to the supplied RNeasy Mini Handbook which contains a complete protocol for sample disruption and RNA purification Important points before starting E Before beginning the procedure read Important Notes page 13 E Ensure that you are familiar with operating the TissueLyser by referring to the operating instructions E If using the RNeasy Mini Kit for RNA purification read the supplied handbook carefully before starting Procedure 1 Pellet the yeast cells by centrifugation Immediately add the appropriate volum
26. nd DNA purification Important points before starting E Before beginning the procedure read Important Notes page 13 E Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions E f using the DNeasy Plant Maxi Kit for DNA purification read the supplied handbook carefully before starting M Fresh frozen or lyophilized tissues can be processed Fresh tissues can be disrupted in lysis buffer at ambient temperature Alternatively fresh or frozen tissues can be disrupted without lysis buffer if the jar containing the sample is frozen in liquid nitrogen Lyophilized tissues can be disrupted without lysis buffer at ambient temperature Disruption of tissues in lysis buffer yields DNA ideal for PCR while disruption of tissues frozen in liquid nitrogen yields DNA of a higher molecular weight We do not recommend disrupting frozen tissues in lysis buffer as this results in low yields and degraded DNA Procedure l Place the tissues in 10 ml grinding jars containing 1 stainless steel grinding ball 20 mm mean diameter 2 f purifying DNA of higher molecular weight from fresh or frozen tissues freeze the jars in liquid nitrogen for 1 min The grinding jars do not need to be frozen if disrupting fresh tissues in lysis buffer or if disrupting lyophilized tissues 3 If necessary add an appropriate volume of lysis buffer e g Buffer AP1 to each jar Lysis buffer must not be added if processin
27. nds Orders 0800 0229592 Fax 0800 0229593 Technical 0800 0229602 Norway Orders 800 18859 Fax 800 18817 Technical 800 18712 Singapore Orders 1800 742 4362 Fax 65 6854 8184 Technical 1800 742 4368 Spain Orders 91 630 7050 Fax 911 630 5145 Technical 91 630 7050 Sweden Orders 020 790282 Fax 020 790582 Technical 020 798328 Switzerland Orders 055 254 22 11 Fax 055 254 22 13 Technical 055 254 22 12 UK Orders 01293 422 911 Fax 01293 422 922 Technical 01293 422 999 USA Orders 800 426 8157 Fax 800 718 2056 Technical 800 DNA PREP 800 362 7737 QIAGEN eee Sample amp Assay Technologies
28. of DNA RNA and protein from cells and tissues AllPrep DNA RNA For 50 preps AllPrep DNA Spin 80004 Protein Mini Kit 50 Columns RNeasy Spin Columns Collection Tubes Buffers AllPrep DNA RNA Kits for simultaneous purification of DNA and RNA from cells and tissues AllPrep DNA RNA For 50 preps AllPrep DNA Spin 80284 Micro Kit 50 Columns RNeasy MinElute Spin Columns Collection Tubes Carrier RNA Buffers AllPrep DNA RNA For 50 preps AllPrep DNA Spin 80204 Mini Kit 50 Columns RNeasy Spin Columns Collection Tubes Buffers GlAamp DNA Mini Kit for purification of genomic mitochondrial bacterial parasite or viral DNA GlAamp DNA Mini Kit 50 For 50 preps GlAamp Spin Columns 51304 Collection Tubes Proteinase K Buffers 34 Tissuelyser Handbook 10 2010 Ordering Information Product Contents Cat no EZ1 DNA Tissue Kit for automated purification of genomic DNA from 1 6 human samples on the BioRobot EZ1 workstation EZ1 DNA Tissue Kit 48 For 48 preps Reagent Cartridges 953034 Tips Tip Holders Tubes Proteinase K Buffer G2 MagAttract DNA Mini M48 Kit for automated purification of genomic DNA from 6 48 human samples on the BioRobot M48 workstation MagAttract DNA Mini For 192 preps MagAttract 953336 M48 Kit 192 Suspension B Proteinase K Buffers GlAsymphony DNA Kits for purification of DNA from a wide range of sample types on the GlAsymphony SP GlAsymphony DNA For 96 preps of 400 pl each 9
29. on and homogenization depends on the tissue being processed and can be extended until no tissue debris is visible Rearranging the tubes ensures uniform disruption and homogenization If processing fiber rich tissues complete disruption and homogenization may sometimes not be possible However small amounts of debris have no effect on subsequent RNA purification with QIAGEN kits and are usually digested in the proteinase K step 5 Proceed with RNA DNA RNA or DNA RNA protein purification Do not reuse the stainless steel beads 5 253 8 an aF Z5 2a Tissuelyser Handbook 10 2010 Protocol Purification of RNA from Plant Tissues This protocol provides guidelines on disrupting plant tissues for subsequent RNA purification If using a QIAGEN kit for RNA purification see Table 3 page 9 refer to the supplied handbook which contains a complete protocol for sample disruption and RNA purification Important points before starting E Before beginning the procedure read Important Notes page 13 E Ensure that you are familiar with operating the Tissuelyser by referring to the operating instructions E Ifusing a QIAGEN kit for RNA purification read the supplied handbook carefully before starting wn oO 2 Sx Z r3 EO 8 E Soft fresh tissues from plants such as Nicotiana and Arabidopsis can often be disrupted and homogenized in lysis buffer Hard tissues e g woody plant materials may require freezing
30. pin columns RNeasy Plus Mini Up to 30 mg tissue includes gDNA 16 Kit Eliminator spin columns automatable on GlAcube Fiber rich RNeasy Fibrous Up to 30 mg tissue 16 tissues e g Tissue Mini Kit heart and RNeasy Fibrous Up to 250 mg tissue 16 muscle Tissue Midi Kit Any type of RNeasy Lipid Tissue Up to 100 mg tissue automatable 16 tissue Mini Kit on GlAcube including miRNeasy Mini Kit Up to 100 mg tissue automatable 16 fatty tissues on GlAcube e g adipose tissue and brain Tissuelyser Handbook 10 2010 7 Table 2 Kits for RNA purification from animal or human tissues using magnetic particles or 96 well plates Sample type Kit Kit format Page Easy to lyse EZ1 RNA Tissue Magnetic particles up to 10 mg tissue 16 tissues e g Mini Kit automated on EZ1 Advanced kidney 1 6 samples per run liver and MagAttract RNA Magnetic particles up to 10 mg tissue 16 lung Tissue Mini M48 automated on BioRobot M48 Kit 6 48 samples per run GlAsymphony Magnetic particles up to 50 mg tissue 16 RNA Kit automated on QlAsymphony SP 1 96 samples per run Any type of EZ1 RNA Universal Magnetic particles up to 50 mg tissue 16 tissue Tissue Kit automated on EZ1 Advanced 1 6 samples per run MagAttract RNA Magnetic particles up to 50 mg tissue 16 Universal Tissue automated on BioRobot M48 M48 Kit 6 48 samples per run RNeasy 96 96 well plate up to 100 mg tissue 16 Universal Tissue Kits automatable on BioRobot Un
31. propylene tubes 19560 1 2 ml 960 in racks of 96 Collection Microtube Caps Nonsterile polypropylene caps for 19566 collection microtubes 1 2 ml and round well blocks 960 in strips of 8 Tissuelyser Handbook 10 2010 31 Ordering Information Product Contents Cat no Related products RNeasy Kits for purification of total RNA from cells tissues and yeast RNeasy Micro Kit 50 For 50 preps RNeasy MinElute 74004 Spin Columns Collection Tubes DNase Carrier RNA Buffers RNeasy Mini Kit 50 For 50 preps RNeasy Spin Columns 74104 Collection Tubes Buffers RNeasy Protect Kits for stabilization and purification of total RNA from tissues RNeasy Protect Mini Kit 50 For 50 preps RNAlater RNA 74124 Stabilization Reagent RNeasy Spin Columns Collection Tubes Buffers RNeasy Plus Kits for purification of total RNA from cells and tissues using gDNA Eliminator spin columns RNeasy Plus Micro Kit 50 For 50 preps RNeasy MinElute Spin 74034 Columns gDNA Eliminator Spin Columns Collection Tubes Carrier RNA Buffers RNeasy Plus Mini Kit 50 For 50 preps RNeasy Spin Columns 74134 gDNA Eliminator Spin Columns Collection Tubes Buffers Reagent DX 1 ml Reagent DX in a screw cap tube 19088 RNeasy Fibrous Tissue Kits for purification of total RNA from fiber rich tissues RNeasy Fibrous Tissue For 50 preps RNeasy Spin Columns 74704 Mini Kit 50 Collection Tubes Proteinase K DNase Buffers RNeasy Fibrous Tiss
32. r to the operating instructions supplied with the Tissuelyser Disruption is carried out in high speed 20 30 Hz shaking steps Disruption for 2 x 3 minutes at 20 30 Hz is usually sufficient to release RNA If disrupting samples for subsequent DNA purification disruption times should be shorter in order to prevent DNA shearing When using a Tissuelyser Adapter Set samples nearer to the Tissuelyser move more slowly than samples further away from the Tissuelyser To ensure uniform disruption and homogenization 2 shaking steps should be carried out After the first shaking step the TissueLyser Adapter Set should be disassembled and the rack of tubes should be rotated so that the tubes that were nearest to the Tissuelyser are now outermost The Tissuelyser Adapter Set should then be reassembled before continuing with the second shaking step For optimal operation the TissueLyser should always be balanced A balance can be provided by assembling a second Tissuelyser Adaptor Set with a rack of tubes containing only disruption beads and fixing this adaptor set into the empty clamp If using grinding jars the balance should consist of a second grinding jar containing a grinding ball Disruption and homogenization in Buffer RLT Plus RNeasy Plus Kits and certain AllPrep Kits are supplied with Buffer RLT Plus a lysis buffer that provides optimal sample lysis as well as appropriate conditions for DNA binding to gDNA Eliminator columns or AllPrep DNA
33. re Kit for manual or automated purification of total DNA from plants in 96 well format MagAttract 96 DNA Plant For 6 x 96 preps MagAttract 67161 Core Kit 6 Suspension A RNase A Buffers BioSprint 15 DNA Plant Kit for automated purification of total DNA from plant tissue on the BioSprint 15 workstation BioSprint 15 DNA Plant Kit 60 For 60 preps MagAttract Suspension 941514 G Rod Covers Tube Strips RNase A Buffers BioSprint 96 DNA Plant Kit for automated purification of total DNA from plant tissue on the BioSprint 96 workstation BioSprint 96 DNA For 576 preps MagAttract 941557 Plant Kit 576 Suspension G Rod Covers Microplates MP S Blocks RNase A Buffer RPW For up to date licensing information and productspecific disclaimers see the respective QIAGEN kit handbook or user manual QIAGEN kit handbooks and user manuals are available at www giagen com or can be requested from QIAGEN Technical Services or your local distributor 36 Tissuelyser Handbook 10 2010 Notes Tissuelyser Handbook 10 2010 37 Notes 38 Tissuelyser Handbook 10 2010 Trademarks QIAGEN GlAamp QlAcube GlAsymphony QlAzol AllPrep BioRobot BioSprint DNeasy EZ1 MagAttract MinElute RNAprotect RNeasy TissueRuptor QIAGEN Group Eppendorf Eppendorf AG Teflon E du Pont de Nemours and Company RNA ater is a trademark of AMBION Inc Austin Texas and is covered by various U S and foreign p
34. s bead dispenser dispenses 96 beads 5 mm diameter in parallel into Collection Microtubes racked enabling high throughput disruption and homogenization The reservoir holds approximately 300 beads The dispenser can be cleaned with water or detergent For more information see the product sheet supplied with the Tissuelyser Bead Dispenser 96 well Grinding Jar Set S Steel The grinding jars allow disruption of 2 large samples in parallel using stainless steel grinding balls Sample disruption can be carried out at room temperature or after freezing the grinding jars in liquid nitrogen For more information see the product sheet supplied with the Grinding Jar Set Grinding Jar Set Teflon The grinding jars allow disruption of 2 large samples in parallel using Teflon grinding balls Sample disruption can be carried out at room temperature For more information see the product sheet supplied with the Grinding Jar Set 28 Tissuelyser Handbook 10 2010 Appendix B Automated Solutions Automated purification using QIAGEN spin column kits Purification of genomic DNA or total RNA from tissues can be fully automated on the QlAcube The innovative QlAcube uses advanced technology to process QIAGEN spin columns enabling seamless integration of automated low throughput sample prep into your laboratory workflow Sample preparation using the QlAcube follows the same steps as the manual procedure i e lyse bind wash and elute enabling you to
35. s for yeast and unicellular animal cells and 3 7 mm stainless steel or tungsten carbide beads for plant and animal human tissues It is essential that glass beads are pretreated before use by washing in concentrated nitric acid Pretreated acid washed beads can be purchased from many vendors of biological supplies e g Sigma cat nos G1145 G1277 and G8772 Disruption parameters for samples not addressed in this handbook must be determined empirically For disruption of large samples a Grinding Jar Set can be used which is supplied with either stainless steel grinding balls for disrupting hard samples such as bone or for disrupting samples in liquid nitrogen or Teflon grinding balls for disrupting most samples Note Do not use Buffer RLT Buffer RLT Plus or GlAzol Lysis Reagent in conjunction with tungsten carbide beads These buffers react chemically with tungsten carbide causing damage to the bead surface When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate material safety data sheets MSDSs available from the product supplier This is not a complete list of suppliers and does not include many important vendors of biological supplies 14 Tissuelyser Handbook 10 2010 Operating the TissueLyser The Tissuelyser Adapter Set or Grinding Jar Set should be securely fixed into the clamps arms of the TissueLyser For details refe
36. s in lysis buffer or if disrupting lyophilized tissues Tissuelyser Handbook 10 2010 23 VNQ uw sanssi JuD d Plant Tissues Mini DNA 4 If necessary add an appropriate volume of lysis buffer e g Buffer AP1 to each tube Lysis buffer must not be added if disrupting precooled tissues or if disrupting lyophilized tissues 5 Place the tubes in the TissueLyser Adapter Set 2 x 24 if using 2 ml tubes or Tissuelyser Adapter Set 2 x 96 if using 1 2 ml tubes 6 Operate the TissueLyser for 1 min at 25 Hz Disassemble the adapter set rotate the rack of tubes so that the tubes nearest to the TissueLyser are now outermost and reassemble the adapter set Operate the TissueLyser for another 1 min at 25 Hz Note If processing precooled tissues increasing the disruption time may lead to thawing and reduced DNA yield and quality 7 Add lysis buffer e g Buffer AP1 if necessary and proceed with DNA purification The tungsten carbide beads can be reused For details on recovering and cleaning beads refer to the DNeasy Plant Handbook 24 Tissuelyser Handbook 10 2010 Protocol Purification of DNA from Plant Tissues Maxi Protocol This protocol provides guidelines on using a Grinding Jar Set to disrupt plant tissues for subsequent DNA purification If using the DNeasy Plant Maxi Kit for DNA purification see Table 5 page 10 refer to the supplied DNeasy Plant Handbook which contains a complete protocol for sample disruption a
37. tion of genomic DNA or total RNA in 96 well System format from animal or human samples plus downstream reaction setup BioSprint 96 Purification of genomic DNA from up to 96 animal or plant samples per run Low throughput sample disruption The TissueRuptor is a handheld rotor stator homogenizer that provides rapid and efficient disruption of individual samples for a wide range of downstream applications The TissueRuptor uses transparent disposable probes which helps to minimize the risk of cross contamination and enables visual control of the sample disruption process The TissueRuptor is an integral part of QIAGEN s complete solution for tissue management in gene expression genotyping and proteomics applications Optimized protocols are available for sample disruption prior to manual or automated nucleic acid or protein purification enabling a streamlined efficient workflow Purification of RNA DNA total nucleic acids or protein can then be performed using QIAGEN kits For more information about the TissueRuptor visit www qiagen com TissueRuptor Automated multicapillary gel electrophoresis The revolutionary QlAxcel System enables fully automated and sensitive high resolution capillary electrophoresis for up to 96 samples per run Ready to go gel cartridges reduce manual handling errors and eliminate the need for tedious gel preparation With the GlAxcel System analysis of DNA fragments single or multiplex PCR produ
38. tocol 23 E Purification of DNA from Plant Tissues Maxi Protocol 25 Appendix A TissueLyser Accessories 27 Appendix B Automated Solutions 29 Ordering Information 31 Tissuelyser Handbook 10 2010 3 Product Contents TissueLyser II Catalog no 85300 Tissuelyser Il 100 120 220 240 V 50 60Hz Operating Instructions Handbook 1 The TissueLyser Il cat no 85300 is an improved version of the Tissuelyser cat no 85200 85210 or 85220 no longer available All instructions and protocols in this handbook apply to both the TissueLyser II and the Tissuelyser Storage The Tissuelyser should be stored upright in a dry environment at room temperature 15 25 C Product Use Limitations The Tissuelyser is intended for molecular biology applications This product is not intended for the diagnosis prevention or treatment of a disease All due care and attention should be exercised in the handling of many of the materials described in this text We recommend all users of QIAGEN products to adhere to the NIH guidelines that have been developed for recombinant DNA experiments or fo other applicable guidelines Product Warranty and Satisfaction Guarantee QIAGEN guarantees the performance of all products in the manner described in our product literature The purchaser must determine the suitability of the product for its particular use Should any product fail to perform satisfactorily due to any reason other th
39. ue For 10 preps RNeasy Spin Columns 75742 Midi Kit 10 Collection Tubes Proteinase K DNase Buffers RNeasy Lipid Tissue Kits for purification of total RNA from all types of tissue including fatty tissues RNeasy Lipid Tissue For 50 preps RNeasy Spin Columns 74804 Mini Kit 50 Collection Tubes QIAzol Lysis Reagent Buffers 32 Tissuelyser Handbook 10 2010 Ordering Information Product Contents Cat no RNeasy 96 Universal Tissue Kits for purification of total RNA from all types of tissue in 96 well format RNeasy 96 Universal For 4 x 96 preps RNeasy 96 Plates 74881 Tissue Kit 4 Elution Microtubes CL Caps S Blocks Airpore Tape Sheets GlAzol Lysis Reagent Buffers RNeasy 96 Universal Tissue For 12 x 96 preps on the BioRobot 967852 8000 Kit 12 Universal System RNeasy 96 Plates Collection Microtubes Elution Microtubes CL Caps S Blocks GlAzol Lysis Reagent Buffers EZ1 RNA Tissue Mini Kit for purification of total RNA from easy to lyse tissues on the BioRobot EZ1 workstation EZ1 RNA Tissue Mini Kit 48 For 48 preps Reagent Cartridges 959034 Tips Tip Holders Tubes DNase Buffer RLT EZ1 RNA Universal Tissue Kit for purification of total RNA from all types of tissue on the BioRobot EZ1 workstation EZ1 RNA Universal For 48 preps Reagent Cartridges 956034 Tissue Kit 48 Tips Tip Holders Tubes QIAzol Lysis Reagent MagAttract RNA Tissue Mini M48 Kit for purification of total RNA from
40. uelyser provides efficient disruption of biological material in each sample vessel for reproducible high quality results in downstream applications such as the purification of total DNA or RNA from a variety of human animal and plant tissues A wide range of GIAGEN sample purification kits are compatible with the Tissuelyser see Tables 1 6 pages 7 10 Sample purification can be performed manually or can be automated using the QlAcube GlAsymphony SP EZ1 Advanced or BioRobof or BioSprint workstations For more information about automated solutions from GIAGEN see Appendix B page 29 6 Tissuelyser Handbook 10 2010 This handbook provides guidelines on disrupting and homogenizing various sample materials for subsequent purification of DNA or RNA Specific details on disruption and homogenization and nucleic acid purification such as the amount of starting material and lysis buffer to use can be found in the handbook supplied with each QIAGEN sample purification kit Table 1 Kits for RNA purification from animal or human tissues using spin columns Sample type Kit Kit format Page Easy tolyse RNeasy Micro Kit Up to 5 mg tissue automatable 16 tissues e g on GlAcube kidney liver RNeasy Mini Kit Up to 30 mg tissue automatable 16 and lung on GlAcube RNeasy Protect Up to 20 mg RNAlater stabilized 16 Mini Kit tissue automatable on GlAcube RNeasy Plus Micro Up to 5 mg tissue includes 16 Kit gDNA Eliminator s
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