FastDNA® SPIN Kit for Soil
Contents
1. Fax 1 440 337 1180 Toll Free Fax 800 334 6999 Germany Singapore MP Biomedicals MP Biomedicals Singapore Europe Phone 0800 426 67337 Tel 65 6775 0008 Toll Free Phone Fax 0800 629 67337 Fax 65 6775 4536 00800 7777 9999 Toll Free Fax 00800 6666 8888 e Switzerland Japan MP Biomedicals Switzerland Australia MP Bio Japan K K Tel 061 271 0007 MP Biomedicals Australasia Tel 03 3808 2102 Fax 061 271 0084 Pty Ltd Toll Free Tel 0120 788 020 Tel 61 2 9838 7422 Fax 03 3808 2401 United Kinad Fax 61 2 9838 7390 ot joi wero MP Biomedicals UK Tel 0800 282 474 i The Netherlands Fax 0800 614 735 Belgium e MP Biomedicals Netherlands MP Biomedicals Tel 02 466 00 00 Hat yang coor Fax 02 466 26 42 Canada Poland MP Biomedicals Canada MP Biomedicals Poland Tel 888 362 5487 Tel 48 22 659 58 95 Fax 514 935 7541 Fax 48 22 658 45 05 www mpbio com MP Biomedicals 29525 Fountain Parkway e Solon OH 44139 tel 1 800 854 0530 fax 1 800 334 69992. Air dry the SPIN Filter for 5 minutes at room temperature 16 Gently resuspend Binding Matrix above the SPIN filter in 50 100 ul of DES DNase Pyrogen Free Water NOTE To avoid over dilution of the purified DNA use the smallest amount of DES required to resuspend Binding Matrix pellet NOTE Yields may be increased by incubation for 5 minutes at 55 C in a heat block or water bath 17 Centrifuge at 14 000 x g for 1 minute to bring eluted DNA into the clean catch tube Discard the SPIN filter DNA is now ready for PCR and other downstream applications Store at 20 C for extended periods or 4 C until use 6 Example Data DNA Isolation from various Soil Samples and Gel Electrophoresis DNA from various soil samples extracted with the FastDNA SPIN kit for Soil 20 of the DNA isolated from 500mg soil was loaded on a 1 2 agarose gel 0 5X TAE Soil was taken from Lane 1 tomato pot Lane 2 sludge Lane 3 sandy soil Lane 4 under pine tree Lane 5 under palm tree Lane 6 green garden Lane 7 Nile Lilly pot Lane 8 lawn grass Lane 9 citrus tree Lane 10 avocado tree DNA ranges from 4 20 kb Fd www mpbio com 1 Recommended Reference Format for Publications DNA was isolated from specific sample using the FastDNA SPIN Kit for Soil and the FastPrep Instrument MP Biomedicals Santa Ana CA 8 References Soil Jacob B lum et al 2006 Appl Envir Microbiol Vol 72 1476 1486 Sedi
3. filters streamline silica handling FastProtein Blue Matrix Cat N 6550 400 50 preps Cat N 6550 500 100 preps Release of proteins from gram positive and gram negative bacteria in 40 seconds Protein extracts are ready for immediate electrophoresis or purification e Ideal for optimizing induction conditions FastProtein Red Matrix Cat N 6550 600 50 preps Cat N 6550 700 100 preps e Release of proteins from yeast cells and fungi in 40 seconds Protein extracts are ready for immediate electrophoresis or purification e Ideal for optimizing induction conditions www mpbio com 13 FastDNA Spin Kit for Soil MP Biomedicals Our family of companies includes f BIO 101 Systems Q 14 www mpbio com Lyse BIG and FAST with Interchangeable Adapters QuickPrep Adapter TeenPrep Adapter 24 x 2 ml samples 12 x 15ml samples ae GER HighPrep Adapter BigPrep Adapter 48 x 2ml samples 2 x 50 ml samples rac The FastPrep 24 instrument is delivered with the QuickPrep Adapter www mpbio com Instruction Manual W FastDNA SPIN Kit for Soil N E Protocol Revision 6560 200 07DEC Worldwide Ordering and Technical Support United States of America France Serbia Worldwide Headquarters MP Biomedicals France MP Global d o o Tel 1 440 337 1200 Tel 03 88 67 54 25 Tel 381 11 2622 945 Toll Free Tel 800 854 0530 Fax 03 88 67 19 45 Fax 381 11 2623 373
4. quick efficient and highly reproducible homogenization that surpasses traditional extraction methods using enzymatic digestion sonication blending douncing and vortexing Samples are placed into 2 0 ml tubes containing Lysing Matrix E a mixture of ceramic and silica particles designed to efficiently lyse all soil organisms including historically difficult sources such as eubacterial spores and endospores gram positive bacteria yeast algae nematodes and fungi Homogenization in the FastPrep Instrument with Lysing Matrix E takes place in the presence of MT Buffer and Sodium Phosphate Buffer reagents carefully developed to protect and solubilize nucleic acids and proteins upon cell lysis These reagents work together to allow extraction of genomic DNA with minimal RNA contamination Following lysis samples are centrifuged to pellet soil cell debris and lysing matrix DNA is purified from the supernatant with a silica based GENECLEAN procedure using SPIN filters Eluted DNA is ready for PCR restriction digest electrophoresis and any other desired application 2 Kit Components and User Supplied Materials 2 1 FastDNA SPIN Kit for Soil Components Lysing Matrix E 50x 2 0 ml tubes Sodium Phosphate Buffer 60 ml MT Buffer 8 ml PPS Solution 25 ml Binding Matrix 66 ml SPIN Modules 50 each Catch Tubes 50 each Concentrated SEWS M 12 ml DES 20ml BBS Gel Loading Dye 200 ul User manual 1 each MSDS 1 each Certificate of Analysis 1 each w
5. Instruction Manual FastDNA SPIN Kit for Soil Rapid Isolation of PCR Ready Genomic DNA from Soil Samples Using the FastPrep System gt One Call _ Size One Source 50 preps storage 2 A W OF d of P a Ambient temperature 15 30 C Biotechnology Reagents Catalog 6560 200 Revision 6560 200 07DEC di www mpbio com MP Biomedicals 29525 Fountain Parkway Solon OH 44139 tel 1 800 854 0530 fax 1 800 334 6999 FastDNA Spin Kit for Soil Lyse BIG and FAST with Interchangeable Adapters QuickPrep Adapter TeenPrep Adapter 24 x 2 ml samples 12 x 15ml samples wwe QI HighPrep Adapter BigPrep Adapter 48 x 2ml samples 2 x 50 ml samples F N The FastPrep 24 instrument is delivered with the QuickPrep Adapter www mpbio com FastDNA SPIN Kit for Soil Rapid Isolation of PCR Ready Genomic DNA from Soil Samples Using the FastPrep System 50 Preps storage Ambient temperature 15 30 C Catalog 6560 200 Revision 6560 200 07DEC www mpbio com FastDNA Spin Kit for Soil TABLE OF CONTENTS 1 Introduction to the FastDNA SPIN Kit for Soil and the FastPrep Instrumenits E U E 5 2 Kit Components and User Supplied Materials 5 2 1 FastDNA SPIN Kit for Soil Componenie 5 2 2 User Supplied Materiales EUEEUEEUUE
6. U 6 3 Important Considerations Before Use 6 3 1 Preparation of SEWS M Wash Solution 6 3 2 Sample Lysis with the FastPrep Instrumeni 6 4 Safety Precautions caccuninacncimasaduneanstannduesedsnunsteciaicendpanintaencneandon 6 O He UO Oh sssrinin iaar niniak tanani 7 6 Example Data DNA Isolation from various Soil Samples and Gel Electrophoresis ccscecescscsssssscseessecsssseseeeerereeseseees 8 7 Recommended Reference Format for Publication 9 8 References ssessssissesesresrsresresrsresrentrtsrnntnrnnnnntnnnnrnnnnnnnnnnnnnnnnn ne 9 9 Related Products ll Klld liukk kk kk kkeke 10 10 Product Use Limitation amp Warraniyi 11 www mpbio com 1 Introduction to the FastDNA SPIN Kit for Soil and the FastPrep Instruments The FastDNA SPIN Kit for Soil quickly and efficiently isolates PCR ready genomic DNA directly from soil samples in less than 30 minutes Designed for use with the FastPrep Instruments from MP Biomedicals plant and animal tissues bacteria algae fungi spores and other members of a soil population are easily lysed within 40 seconds These benchtop devices use a unique optimized motion to homogenize samples by multidirectional simultaneous impaction with lysing matrix particles FastPrep Instruments provide an extremely
7. dits refund of the purchase price of or the replacement of materials that do not meet our specification By acceptance of the product Buyer indemnifies and holds MP Biomedicals harmless against and assumes all liability for the consequence of its use or misuse by the Buyer its employees or others including but not limited to the cost of handling Said refund or replacement is conditioned on Buyer notifying MP Biomedicals Inc within thirty 30 days of receipt of product Failure of Buyer to give said notice within thirty 30 days shall constitute a waiver by the Buyer of all claims hereunder with respect to said material s FastDNA FastRNA FastPrep and BIO 101 Systems are registered trademarks of MP Biomedicals LLC www mpbio com 11 FastDNA Spin Kit for Soil Ready to use Protocols For DNA RNA And Protein Isolation From Any Sample Rapid and reproducible sample lysis and purification process e No cross contamination with the closed lysing matrix tubes Increased yields of high quality DNA RNA and proteins Integrity and size of DNA RNA and proteins are retained e Nucleic acids and proteins are ready to use in downstream application DNA PROTEIN FastPrep Kits RNA FastDNA Kit and FastDNA Spin Kit Cat N 6540 400 Cat N 6540 600 respectively 100 preps Lyse and isolate DNA in less than 30 minutes Plant animal yeast fungal and microbial samples No hazardous organic reage
8. itation Solution and mix by shaking the tube by hand 10 times 7 Centrifuge at 14 000 x g for 5 minutes to pellet precipitate Transfer supernatant to a clean 15 ml tube NOTE While a 2 0 ml microcentrifuge tube may be used at this step better mixing and DNA binding will occur in a larger tube 8 Resuspend Binding Matrix suspension and add 1 0 ml to supernatant in 15 ml tube 9 Place on rotator or invert by hand for 2 minutes to allow binding of DNA Place tube in a rack for 3 minutes to allow settling of silica matrix 10 Remove and discard 500 ul of supernatant being careful to avoid settled Binding Matrix 11 Resuspend Binding Matrix in the remaining amount of supernatant Transfer approximately 600 ul of the mixture to a SPIN Filter and centrifuge at 14 000 x g for 1 minute Empty the catch tube and add the remaining mixture to the SPIN Filter and centrifuge as before Empty the catch tube again 12 Add 500 ul prepared SEWS M and gently resuspend the pellet using the force of the liquid from the pipet tip NOTE Ensure that ethanol has been added to the Concentrated SEWS M See section 3 1 www mpbio com 7 FastDNA Spin Kit for Soil 13 Centrifuge at 14 000 x g for 1 minute Empty the catch tube and replace 14 Without any addition of liquid centrifuge a second time at 14 000 x g for 2 minutes to dry the matrix of residual wash solution Discard the catch tube and replace with a new clean catch tube 15
9. ment Tracy J Mincer et al 2005 Appl Envir Microbiol Vol 71 7019 7028 Snow samples Takahiro Segawa et al 2005 Appl Envir Microbiol Vol 71 123 130 Feces Alice Layton et al 2006 Appl Envir Microbiol Vol 72 4214 4224 sediments and Soil Francis C A et al 2005 PNAS Vol 102 14683 14688 Soil Akira Ando et al 2005 Appl Envir Microbiol Vol 71 7075 7082 www mpbio com 9 FastDNA Spin Kit for Soil 9 Related Products Description Siz Catalog FastPrep 24 Instrument 6002 500 FastPrep FP100A Instrument 6001 100 FastPrep FP120A Instrument 6001 120 FastPrep FP220A Instrument 6001 220 FastDNA Kit 6540 400 FastDNA SPIN Kit 6540 600 FastRNA Pro Red Kit Yeast amp Fungus FastRNA Pro Green Kit Plant amp Animal www mpbio com 10 Product Use Limitation amp Warranty Unless otherwise indicated this product is for research use only Purchase of MP Biomedicals products does not grant rights to reproduce modify or repackage the products or any derivative thereof to third parties MP Biomedicals makes no warranty of any kind expressed or implied including merchantability or fitness for any particular purpose except that the products sold will meet our specifications at the time of delivery Buyer s exclusive remedy and the sole liability of MP Biomedicals hereunder shall be limited to at our discretion no replacement or compensation product cre
10. nts required e SPIN filters streamline silica handling FastDNA Spin Kit FastDNA Spin Kit for Soil Cat N 6560 200 100 preps e Lyse and isolate DNA in less than 30 minutes Variety of soil and environmental sample types No hazardous organic reagents required SPIN filters streamline silica handling 12 www mpbio com FastRNA Pro Blue Kit Cat N 6025 050 50 preps For use with gram positive and gram negative bacteria e Lyse up to 10 cells per 2ml tube e Lysis and isolation with single phase organic solution in less than 90 minutes FastRNA Pro Red Kit Cat N 6035 050 50 preps For use with yeast cells and fungal tissue e Lyse up to 10 cells per 2ml tube e Lysis and isolation with single phase organic solution in less than 90 minutes FastRNA Pro Green Kit Cat N 6045 050 50 preps For use with all plant and animal samples Lyse 50 100 mg tissue per 2ml tube e Lysis and isolation with single phase organic solution in less than 90 minutes FastRNA Pro Soil Direct Kit and FastRNA Pro Soil Indirect Kit Cat N 6070 050 Cat N 6075 050 respectively 50 preps e Isolate RNA from soil samples direct kit and washed soil indirect kit in less than 2 hours Variety of soil and environmental sample types RNA protected during and after processing Humic acids reduced to allow uninhibited RT PCR Includes additional reagents for even further purification if necessary SPIN
11. ther products with the FastPrep Instrument is not recommended and may result in sample loss or instrument failure A single 40 second run at a speed setting of 6 0 in the FastPrep Instrument is sufficient to lyse almost all samples If the user experimentally determines that additional processing time is required the sample should be incubated on ice in the Lysing Matrix E tube for at least 2 minutes between successive FastPrep Instrument homogenizations to prevent overheating the sample and tube 4 Safety Precautions Binding Matrix contains components that when in contact with human tissue may cause irritation Wear personal protective equipment to prevent contact with the skin or mucus membranes gloves lab coat and eye protection Consult the enclosed Material Safety Data Sheet for additional details 6 www mpbio com 5 Protocol 1 Add up to 500 mg of soil sample to a Lysing Matrix E tube NOTE See section 3 2 for important guidelines 2 Add 978 ul Sodium Phosphate Buffer to sample in Lysing Matrix E tube 3 Add 122 ul MT Buffer 4 Homogenize in the FastPrep Instrument for 40 seconds at a speed setting of 6 0 5 Centrifuge at 14 000 x g for 5 10 minutes to pellet debris NOTE Extending centrifugation to 15 minutes can enhance elimination of excessive debris from large samples or from cells with complex cell walls 6 Transfer supernatant to a clean 2 0 ml microcentrifuge tube Add 250 ul PPS Protein Precip
12. ww mpbio com 5 FastDNA Spin Kit for Soil 2 2 User Supplied Materials FastPrep Instrument See Section 9 Microcentrifuge that can freely spin 2 0 ml tubes Microcentrifuge tubes 2 0 ml and 1 5 ml Clean 15 ml tubes for DNA binding Rotator or low speed vortex 3 Important Considerations Before Use 3 1 Preparation of SEWS M Wash Solution The FastDNA SPIN Kit for Soil contains a bottle with 12 ml of Concentrated SEWS M Wash Solution Before using this solution add 100 ml of 100 ethanol and mark on the bottle label the date ethanol was added Ensure that the bottle is securely closed to prevent evaporation mix and store at room temperature 3 2 Sample Lysis with the FastPrep Instrument The fill volume in the lysing matrix tube after the addition of the Sodium Phosphate and MT Buffers to the sample should allow sufficient air soace in the sample tube for efficient FastPrep Instrument processing MP Biomedicals recommends using 500 mg of starting material as long as there is between 250 500 ul of empty space in the tube Sample loss or tube failure may result from overfilling the matrix tube The matrix tube caps must be secure but not over tightened to prevent sample leakage If the sample is too large for processing in a single tube divide the sample and process using multiple tubes MP Biomedicals Lysing Matrix particles and tubes have been rigorously tested and validated in the FastPrep Instrument The use of o
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