FastDNA SPIN kit
Contents
1. Binding Matrix Invert to mix NOTE It is important to use a tube that is large enough to allow room for complete mixing of the entire volume during the course of the next step Tubes with conical bottoms are not recommended A 2 0 ml microcentrifuge tube will work well at this step 6 Incubate with gentle agitation for 5 minutes at room temperature on a rotator NOTE A low speed vortex may be used at this point but care must be taken not to shear the DNA 7 Centrifuge at 14 000 x g for 10 seconds to pellet Binding Matrix Discard supernatant 8 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 9 Transfer the resuspended Binding Matrix to a SPIN Module SPIN filter and Catch Tube Centrifuge at 14 000 x g for 1 minute Discard contents of Catch Tube and replace 10 Centrifuge a second time at 14 000 x g for 1 minute and replace the Catch Tube with a Recovery Tube 11 Elute DNA by gently resuspending Binding Matrix above the SPIN Filter in 100 ul of DES Incubate for 5 minutes at 55 C in a heat block or water bath 12 Centrifuge at 14 000 x g for 1 minute to bring eluted DNA into the Recovery Tube Discard the SPIN Filter DNA is now ready for downstream applications Store at 20 C for extended periods or 4 C until use 6 Recommended Reference Format for Publications DNA was isolat2. Recommended Reference Format for Publication 8 7 References 4 aaa 8 8 Related Products a 9 9 Product Use Limitation amp Warranty 000 cece eee eee eee 9 4 visit us on the web at www qbiogene com FastDNA SPIN Kit 1 Introduction to the FastDNA SPIN Kit and the FastPrep Instrument The FastDNA SPIN Kit quickly and efficiently isolates genomic DNA from a wide variety of sources Designed for use with the FastPrep Instrument plant and animal tissues bacteria algae fungi and many other sam ples are easily lysed within 40 seconds This benchtop device uses a patented vertical angular motion to homogenize samples by multidirectional simultaneous impaction with lysing matrix particles The FastPrep Instrument provides an extremely quick efficient and highly reproducible homogenization that surpasses tra ditional extraction methods using enzymatic digestion sonication blending douncing and vortexing Samples are placed into 2 0 ml tubes containing Lysing Matrix A irregularly shaped garnet particles and a single 1 4 inch ceramic sphere While almost all samples are easily processed with this pre filled combi nation additional 1 4 inch ceramic spheres are provided for hard samples such as bone cartilage or seeds Homogenization in the FastPrep Instrument with Lysing Matrix A takes place in the presence of sample spe cific Cell Lysis Solutions CLS For plant tissues CLS VF is
3. used in conjunction with a Protein Precipitation Solution PPS Yeast algae and fungi are lysed in the presence of CLS Y For all other samples CLS TC is used during sample lysis For maximum flexibility all buffers are provided in the kit Following lysis samples are centrifuged to pellet debris and lysing matrix DNA is purified from the super natant with a silica based GENECLEAN procedure using SPIN filters Eluted DNA is ready for digestion electrophoresis PCR and any other desired application 2 Kit Components and User Supplied Materials 2 1 FastDNA SPIN Kit Components Lysing Matrix A 100 2 0 ml tubes 1 4 Ceramic Spheres 100 spheres Binding Matrix 66 ml Concentrated SEWS M 12 ml DES 25 ml CLS VF 90 ml PPS 25 ml CLS TC 110 ml CLS Y 110 ml SPIN Modules 100 each Recovery Tubes 100 each User manual 1 each MSDS 1 each Certificate of Analysis 1 each Any Questions Call Technical Support at 800 424 6101 5 FastDNA SPIN Ki 2 2 User Supplied Materials FastPrep Instrument Cat 6001 100 120 or 220 Microcentrifuge that can freely spin 2 0 ml tubes Microcentrifuge tubes 2 0 ml and 1 5 ml Rotator or low speed vortex 3 Important Considerations Before Use 3 1 Preparation of SEWS M Wash Solution The FastDNA Kit 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 Ensu
4. 00 Phone 0800 328 8401 E mail technical qbiogene com E mail techservuk qbiogene com FRANCE GERMANY Phone 03 88 67 54 25 Phone 0800 409 0580 E mail techservfr qbiogene com E mail techservde qbiogene com www qbiogene com Q BlOg ere
5. Qbiogene Application Manual FastDNA SPIN Kit Rapid Isolation of Genomic DNA from Plant and Animal Tissue Bacteria Yeast Algae and Fungi Using the FastPrep Instrument N Revision 6540 600 4101 Q BlOgene FastDNA SPIN Kit FastDNA SPIN Kit Rapid Isolation of Genomic DNA from Plant and Animal Tissue Bacteria Yeast Algae and Fungi Using the FastPrep Instrument Application Manual Revision 6540 600 4 01 Catalog 6540 600 100 Preps Storage temperature Ambient temperature 15 30 C Any Questions Call Technical Support at 800 424 6101 3 FastDNA SPIN Kit TABLE OF CONTENTS 1 Introduction to the FastDNA SPIN Kit and the FastPrep Instrument 5 2 Kit Components and User Supplied Materials 5 2 FAStDNA SPIN Kit Components 5 2 2 User Supplied Materials s cac2escaaesscaenssanascnconsaanaroinaeassnanavaadessaansivaagvitaeassncadeahansninaaad 6 3 Important Considerations Before Use 000 c cece cee eee eens 6 3 1 Preparation of SEWS M Wash Solution 6 3 2 Precipitate Material in CLS TC Buffer ccc ccccccecsccsscsscsecsscsecsecsessecsesseesassasaseases 6 3 3 Sample Lysis with the FastPrep Instrument ccccccecscsecseseceseesessssecssseestsereseeseees 6 3 4 Recovery of DNA from Dry Samples 7 3 00 GO PUNMGATION OF RNA sssaaa EEA RANE 7 4 Safty PRECAUTIONS ido 7 5 PHOTO GON carinae a a nes a a a a E nce eantesei 7 6
6. arch use only Purchase of Qbiogene Inc products does not grant rights to reproduce modify or repackage the products or any derivative thereof to third parties Qbiogene Inc 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 Qbiogene Inc hereunder shall be limited to at our dis cretion no replacement or compensation product credits refund of the purchase price of or the replace ment of materials that do not meet our specification By acceptance of the product Buyer indemnifies and holds Qbiogene Inc 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 Qbiogene 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 Qbiogene Inc Any Questions Call Technical Support at 800 424 6101 9 FastDNA SPIN Kit NOTES 10 visit us on the web at www gbiogene com NORTH AMERICA UK Phone 1 800 424 6101 or 760 929 17
7. ed from specific sample using the FastDNA SPIN Kit and the FastPrep Instrument Qbiogene Inc CA 7 References A wide variety of references for lysis and purification with the FastPrep products can be found on our web site at http www qbiogene com fastprep FastPrepDB index shtml 8 visit us on the web at www qbiogene com FastDNA SPIN Kit 8 Related Products Item Size Description Catalog FastPrep FP100A Instrument 100V 6001 100 FastPrep FP120A Instrument 120V 6001 120 FastPrep FP220A Instrument 220V 6001 220 FastDNA Kit 100 preps 6540 400 FastDNA SPIN Kit 100 preps 6560 600 FastDNA SPIN Kit for Soil 50 preps 6560 200 FastRNA Pro Soil Direct Kit 50 preps 6070 050 FastRNA Pro Soil Indirect Kit 50 preps 6075 050 FastRNA Pro Red Kit Yeast 50 preps 6035 050 FastRNA Pro Green Kit Plant amp Animal 50 preps 6045 050 FastRNA Pro Blue Kit Bacteria 50 preps 6025 050 FastProtein Blue Matrix 50 preps 6550 400 FastProtein Red Matrix 50 preps 6550 600 Lysing Matrix A 50 x 2 ml tubes 6910 050 Lysing Matrix A 100 x 2 ml tubes 6910 100 Lysing Matrix A 50 x 2 ml tubes 6910 500 QA Agarose Molecular Biology Grade 500 g AGAHO500 BBG general purpose neutral gel RNA and DNA loading dye with glycerol 1 ml 2327 104 BBS general purpose neutral gel RNA and DNA loading dye with sucrose 1 ml 2325 104 9 Product Use Limitation amp Warranty Unless otherwise indicated this product is for rese
8. les If the user experimentally determines that additional processing time is required the sample should be incubated on ice in the Lysing Matrix A tube for at least 2 minutes between successive FastPrep Instrument homogenizations to prevent overheating the sample and tube Qbiogene recommends that all researchers begin the protocol with the Lysing Matrix A as supplied in the kit Garnet matrix and single sphere If lysis is inefficient even after multiple runs of 40 seconds an addi tional 1 4 inch ceramic sphere can be added on top of the sample Depending on the sample lysis and or 6 visit us on the web at www qbiogene com FastDNA SPIN Kit yield may or may not improve and shearing of existing genomic DNA may begin to occur Samples with 2 spheres should be processed carefully in order to balance increased yield and lysis against increased DNA shearing by varying speed and or time settings 3 4 Recovery of DNA from Dry Samples To optimize DNA recovery from extremely dry samples leave the lysed sample at room temperature in the Lysing Matrix A tube for an incubation period of 15 minutes to 2 hours after processing in the FastPrep Instrument 3 5 Co Purification of RNA Some tissues i e liver kidney contain very high levels of RNA which may co purify with the genomic DNA If absolute control of RNA contamination is necessary the final eluted DNA can be treated with RNase as per the manufacturer s protocol 4 Safety Preca
9. re that the bottle is securely closed to prevent evaporation and store at room temperature 3 2 Precipitate Material in CLS TC Buffer If the FastDNA Kit was shipped or stored at a low temperature a harmless precipitate may form in the CLS TC Buffer If a precipitate is seen incubate the bottle in a 45 55 C water bath for several minutes and mix to bring the precipitate back into solution Allow solution to cool to room temperature 3 3 Sample Lysis with the FastPrep Instrument The fill volume in the lysing matrix tube after the addition of the Cell Lysis Solution to the sample should allow sufficient air space in the sample tube for efficient FastPrep Instrument processing Qbiogene rec ommends using 100 200 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 pro cessing in a single tube divide the sample and process using multiple tubes Qbiogene s Lysing Matrix particles and tubes have been rigorously tested and validated in the FastPrep Instrument The use of non Qbiogene 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 samp
10. utions 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 5 Protocol 1 Add sample to Lysing Matrix A tube Place up to 100 200 mg tissue fresh frozen dried etc or 200 ul of cells suspended in water or isotonic saline solution For bacteria yeast algae or tissue culture cells grown in suspension Centrifuge a sufficient volume of culture to provide a pellet size of 50 100 mg wet weight or up to 109 bacteria 108 yeast algae or 107 mammalian cells Resuspend pellets in water or isotonic saline to give a maximum suspension volume of 200 ul NOTE See section 3 3 for other important guidelines 2 Add appropriate Cell Lysis Solution CLS according to table below Processing Tissue From Add to Sample Tube Plant tissue 800 ul CLS VF and 200 ul PPS Animal tissue cultured cells insects bacteria bone etc 1 0 ml CLS TC Yeast algae or fungi 1 0 ml CLS Y 3 Homogenize in the FastPrep Instrument for 40 seconds at a speed setting of 6 0 4 Centrifuge at 14 000 x g for 5 10 minutes to pellet debris Any Questions Call Technical Support at 800 424 6101 7 FastDNA SPIN Kit 5 Transfer supernatant 600 700 ul to a 2 0 ml microcentrifuge tube and add an equal volume of
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