UltraClean Standard Mini Plasmid Prep Catalog No. Quantity 12301-50 50 Preps 12301-100 100 Preps 12301-250 250 Preps Instruction Manual New & Improved Please read for protocol changes! F Please recycle Version: 10292010 1
Table of Contents Introduction... 3 Protocol Overview... 3 Flow Chart... 4 Equipment Required... 5 Contents & Storage... 5 Precautions & Warnings... 5 Protocols: Detailed Protocol (Describes what is happening at each step)... 6 Short Protocol... 9 Vacuum Manifold Protocol...10 Hints & Troubleshooting Guide...12 Contact Information...14 Other Quality Products Available...15 2
Introduction The UltraClean Standard Mini Plasmid Prep is based on the standard alkaline lysis technique used in most plasmid prep protocols. The kit uses the same reagents and volumes as in most competitor kits so you do not need to learn a new protocol. Our optimized reagents and shorter centrifuge times provide a protocol that is quicker and easier than others. Its silica spin filters purify and recover the highest possible yields of sequence quality plasmid. Compatible with LB and all high nutrient growth media, the UltraClean Standard Mini Plasmid Prep produces plasmids free of genomic DNA contamination. The DNA is concentrated enough to proceed directly with automated sequencing and can also be used in restriction digests, ligation, PCR and other downstream applications. Protocol Overview This kit will purify plasmid DNA from 1-5 ml of bacterial culture. Cells are lysed with an improved set of alkaline lysis reagents. Plasmid DNA is bound to a silica spin filter, washed once, and recovered in Tris buffer or water. The DNA is concentrated enough to proceed directly with any of the downstream applications listed in the introduction above. High Throughput Options MO BIO offers a vacuum based protocol for faster processing without centrifugation for the DNA binding and column washing steps for Spin Filters. The MO BIO PowerVac Manifold allows for processing of up to 20 spin filter preps at a time using the PowerVac Mini Spin Filter Adapters. For additional high throughput options MO BIO offers the UltraClean -htp 96 Well Plasmid Prep for processing up to 2 x 96 samples using a centrifuge capable of spinning two 96 Well Blocks stacked (13 cm x 8 cm x 5.5 cm) at 4500 x g. For 96 well homogenization MO BIO offers the 96 Well Plate Shaker and Plate Adapter Set (MO BIO Catalog# 11996 & 11999, respectively.) This kit is for research purposes only. Not for diagnostic use. Other Related Products Catalog No. Quantity TB DRY Powder Growth Media 12105-05 12105-1 12105-5 12105-10 12105-20 10 kg 20 kg UltraClean 6 Minute Mini Plasmid Prep 12300-50 12300-100 12300-250 2 UltraClean -htp 96 Well Plasmid Prep 12396-4 12396-12 4 x 96 preps 12 x 96 preps PowerVac Manifold 11991 1 manifold PowerVac Mini System 11992 1 unit + 20 adapters PowerVac Mini Spin Filter Adapters 11992-10 11992-20 10 adapters 20 adapters 3
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Equipment Required Microcentrifuge (13,000 x g) Vortex-Genie 2 Vortex (MO BIO Catalog# 13111-V or 13111-V-220) 80 well microcentrifuge tube rack (optional) Pipettors: 50 700µl Reagents Required but not Included 100% ethanol (for the PowerVac Manifold protocol only) Contents Catalog # 12301-50 Catalog # 12301-100 Catalog # 12301-250 Component Catalog # Amount Catalog # Amount Catalog # Amount Solution 1 12301-50-1 5.5 ml 12301-100-1 11 ml 12301-250-1 28 ml Solution 2 12301-50-2 11 ml 12301-100-2 22 ml 12301-250-2 55 ml Solution 3 12301-50-3 22 ml 12301-100-3 44 ml 12301-250-3 110 ml Solution 4 12301-50-4 30 ml 12301-100-4 2 x 30 ml 12301-250-4* 83 ml* Solution 5 12301-50-5 3 ml 12301-100-5 6 ml 12301-250-5 15 ml Spin Filters 12301-50-SF 50 12301-100-SF 100 12301-250-SF 250 (Units in 2 ml Tubes) 2 ml Collection 12301-50-T 50 12301-100-T 100 12301-250-T 250 Tubes *NOTE: Before first use catalog# 12301-250-4 (Solution 4) must be prepared. Please see instructions prior to step 1 in the protocol. Storage reagents and components should be stored at room temperature (15-30 C). Precautions Please wear gloves when using this product. Avoid all skin contact with kit reagents. In case of contact, wash thoroughly with water. Do not ingest. See Material Safety Data Sheets for emergency procedures in case of accidental ingestion or contact. All MSDS information is available upon request (760-929- 9911) or at www.mobio.com. Reagents labeled flammable should be kept away from open flames and sparks. WARNING: Solution 4 is flammable. Avoid contact of Solution 3 with bleach or other oxidizers. Do not use bleach to clean the inside of the PowerVac Manifold or to rinse the PowerVac Mini Spin Filter Adapters when attached to the manifold.. 5
Detailed Protocol (Describes what is happening at each step) We highly recommend you read this information if this is your first time using the UltraClean Standard Mini Plasmid Prep. Please wear gloves at all times BEFORE THE FIRST USE OF 12301-250-4 ONLY, Solution 4 must be prepared. Add 83 ml of 100% ethanol. Mix well. Put a check mark in the ethanol added box on the bottle cap. 1. Grow cells (plasmid culture) to a typical density of A 600 = 2.0 or higher. 2. Each plasmid prep will require a set of two 2 ml and one Spin Filter unit (a plastic spin basket with a white silica membrane sitting inside a 2 ml Collection Tube). 3. Label the caps of each set with an ethanol resistant lab marker. It is convenient to place these sets in a microcentrifuge tube rack in this order: tube, Spin Filter, 2 ml Collection Tube. 4. Open the first tube in each set and add up to 2 ml of culture. If you are using high nutrient broth (terrific broth, TB DRY, 2 x YT, super broth, etc.), use no more than 2 ml of culture per prep. For LB or low nutrient cultures, you can use 2 ml or you can combine cells from up to 5 ml of culture. Do this by spinning 2 ml of cells, discarding the supernatant, adding more culture to the same tube and spinning again. Repeat until 5 ml worth of cells have been processed. Note: The yields of many low copy plasmids (containing large inserts), can be increased by growing in high nutrient broth such as TB DRY (MO BIO Catalog# 12105-1). 5. Orient the microcentrifuge the same way each time you spin them in all the following procedures so that the tube hinge is facing straight out away from the center of the rotor. 6. Centrifuge for 1 minute at 13,000 x g. What s happening: The bacterial cells are being forced to the bottom of the tube. 7. Decant the supernatant by inverting the tube and pouring into a waste container. You will need to do another spin to be sure to remove all traces of liquid from the sides of the tube as described in the next 3 steps. 8. Orient with the hinges in same position (hinges pointing straight out from center of rotor). 9. Centrifuge 5 seconds at 13,000 x g. 10. Remove all visible liquid with a narrow pipet tip. Removing all liquid at this step is critical. Using a small bore pipet tip helps remove all traces of liquid media. What s happening: The cells have been pelleted and are now separated from the culture growth medium. Removal of trace amounts of growth medium will ensure against dilution of the reagents in all following steps. 11. Add 100 l Solution 1 to each cell pellet tube and close the. 12. Resuspend the bacterial pellet by bump vortexing with the vortex set at the highest speed. Bump vortexing means: hold the tube tip on the vortex head for 10 seconds, take it off for 1 second then hold it on the vortex again. Repeat this process for 1 minute. After 1 minute, hold the tube in a 6
horizontal position up to a light and look at it. The liquid will spread from one end of the tube to the other. If you see any clumps of cells, keep bump vortexing until they are gone. It takes a minimum of 1 minute with the vortex at its highest speed to resuspend cells in the 100 l volume. Do two at a time when processing multiple preps for best efficiency. An alternative procedure is to scrape the tube tips back and forth over the holes of an 80 well microcentrifuge tube rack. This procedure is actually the subject of a paper in Biotechniques. Voo, K. S.; Jacobsen, B. M. BioTechniques 24:240-243, February 1998. What s happening: The bacterial cells are re-suspended in a small volume of buffer that keeps them from breaking open (lysing). It is important to make this suspension of cells homogeneous because cells trapped in clumps will be resistant to lysis reagents. Solution 1 contains RNase A; however, it cannot digest RNA until the cells are lysed in the next step. 13. Check Solution 2. If precipitated, heat to 55-65 C for 5 minutes to dissolve. Mix before use. What s happening: Solution 2 contains a detergent SDS that can precipitate if cooled. This precipitate is easy to re-suspend by heating. For this reason, always store this kit at room temperature (20-25 C). Solution 2 can be used while still warm. 14. Add 200 l Solution 2 to the cell suspension. What s happening: Alkaline cell lysis. Solution 2 is very alkaline (ph 12) and contains the detergent SDS. Addition of Solution 2 causes the bacterial cells to lyse because the proteins in the cell membrane become denatured similar to when you cook an egg. All DNA becomes denatured to its single stranded form at this point. The bacterial chromosomal DNA is long and is attached to broken pieces of the cell membrane. Plasmid DNA is linked so it forms two attached circles. Like two links of a chain. All RNA is digested during this very short step because RNase A is active even in very alkaline conditions. 15. Close all. 16. Gently invert the up to 8 times to mix. Do not vortex at this step. Vortexing will reduce the quality of your plasmid prep. It causes chromosomal DNA contamination by breaking pieces of the bacteria s chromosomal DNA which will then co purify with the plasmid DNA. 17. Add 400 l Solution 3. What s happening: Neutralization. Solution 3 contains potassium acetate and salt. The potassium acetate forms a precipitate when it interacts with SDS. At the same time denatured proteins co-precipitate with the SDS. Solution 3 neutralizes the alkaline ph to a more neutral ph 7. All DNA tries to re-nature. Plasmid can easily re-form to its double stranded form. Bacterial chromosomal DNA finds it difficult to re-nature because it has no reference point and homologous pieces of DNA may be blocked from finding each other by the cell debris present. 18. Close the and gently invert up to 8 times to mix. Do not vortex at this step. Vortexing causes chromosomal DNA contamination. 19. Centrifuge for 3 minutes at 13,000 x g minimum. What s happening: Dense cell debris is pelleted to the bottom of the tube. Chromosomal DNA is also pelleted along with the cell debris. 20. Remove the from the centrifuge. There should be a clear non viscous supernatant on top of a large white pellet stuck to the sides of the tube. If the pellet is not firm but instead it is loose or 7
gloppy, this is a clear indication you did not remove all the culture media when you originally pelleted the cells. You will need to start over in this case and be more careful to remove all the culture media. 21. Open the cap of as many Spin Filters as you have plasmid preps. 22. Transfer all of the clear liquid supernatant to a Spin Filter (avoid the white precipitate). Decanting is the best method here. Just turn the tube so that you pour away from the hinge. The white pellet will stay stuck to the side of the tube. Close the lids of the Spin Filters. 23. Centrifuge 30-60 seconds at 10,000 x g. The liquid will flow through the white spin filter membrane leaving the plasmid DNA bound to the filter membrane. What s happening: The plasmid DNA now binds to the white silica membrane in the spin filter. Plasmid DNA binds due to the high salt conditions. Unwanted impurities such as digested RNA, and any other cell components that did not pellet are passed through the spin filter and end up in the flow through in the collection tube. This flow through is discarded. 24. Lift out the plastic spin filter basket from the 2 ml Collection Tube, discard the liquid from the 2 ml Collection Tube, and then replace the spin filter basket into the Collection Tube. Note: Do not let the liquid discarded from the Spin Filter Collection Tube come in contact with bleach. 25. Add 600 l Solution 4 to the Spin Filter. Close the lid. Note: If you have not added ethanol to 12301-250-4 before first use, do so now as described just before step 1. However, if you are using either 12301-50 or 12301-100, Solution 4 already contains the required volume of ethanol; no additional ethanol needs to be added. 26. Centrifuge 30-60 seconds at 13,000 x g. 27. Discard flow through liquid from the 2 ml Collection Tube, and centrifuge again for 1 minute at 13,000 x g. What s happening: Solution 4 washes the DNA that is bound to the spin filter. Solution 4 is an ethanol wash buffer. The ethanol keeps the plasmid DNA bound to the filter as impurities are washed away. 28. Being careful not to splash liquid on the filter basket, place spin filter basket in a new 2 ml Collection Tube (provided). 29. Add 50 l of Solution 5 or sterile water directly in the middle of the white spin filter membrane. Let stand for 1 minute. What s happening: Solution 5 is 10mM Tris. As it passes through the spin filter, the plasmid DNA is released (eluted) off the filter and it passes into the collection tube. The plasmid DNA is released because it will not stay bound to the spin filter when there is no salt present. 30. Centrifuge 1 minute at 10,000 x g. 31. Remove filter basket and close tube lid. Plasmid DNA in the 2 ml Collection Tube is now ready to use for any application. Thank you for choosing the UltraClean Standard Mini Plasmid Prep! 8
Short Protocol (Use only if you are already familiar with this kit) Note: Remember to orient with hinges pointing straight out from center of centrifuge rotor for all centrifuge steps. Please wear gloves at all times BEFORE THE FIRST USE OF 12301-250-4 ONLY, Solution 4 must be prepared. Add 83 ml of 100% ethanol. Mix well. Put a check mark in the ethanol added box on the bottle cap. 1. Pellet 2 ml of overnight culture by centrifuging for 1 minute at 13,000 x g. Decant liquid media and spin again for 5 seconds. Remove the remaining liquid media with a small bore pipet tip. 2. Resuspend cell pellets in 100 l of Solution 1. Bump vortex until the mixture is completely homogeneous and no clumps exist. 3. Add 200 l of Solution 2 and gently invert up to 8 times to mix. Do not vortex! This will cause shearing of the Genomic DNA. 4. Add 400 l of Solution 3 and gently invert up to 8 times to mix. The resulting solution will be cloudy. 5. Centrifuge for 3 minutes at 13,000 x g. 6. Transfer the supernatant to the Spin Filter unit. Avoid the white pellet. Decanting is the best method to transfer the supernatant. 7. Centrifuge 30 60 seconds at 10,000 x g. Discard the flow through. 8. Add 600 l of Solution 4 to the spin filter unit. 9. Centrifuge for 30 60 seconds at 13,000 x g. 10. Discard the flow through. Important: Centrifuge for 1 additional minute at 13,000 x g to remove the residual wash solution. 11. Transfer the spin filter basket to a 2 ml Collection Tube. Be careful not to splash any Solution 4 on the Spin Filter! 12. Add 50 l of Solution 5 or sterile water to the center of the spin filter membrane. Let stand at room temperature for 1 minute. Centrifuge for 1 minute at 10,000 x g. 13. Remove the spin filter basket, now the DNA is ready for any application. Thank you for choosing the UltraClean Standard Mini Plasmid Prep! 9
Vacuum Protocol using the PowerVac Manifold Please wear gloves at all times For each sample lysate, use one Spin Filter column. Keep the Spin Filter in the attached 2 ml Collection Tube and continue using the Collection Tube as a Spin Filter holder until needed for the Vacuum Manifold Protocol. Label each Collection Tube top and Spin Filter column to maintain sample identity. If the Spin Filter becomes clogged during the vacuum procedure, you can switch to the procedure for purification of the DNA by centrifugation. You will need to provide 100% ethanol for step 4 of this protocol 1. For each prep, attach one aluminum PowerVac Mini Spin Filter Adapter (MO BIO Catalog# 11992-10 or 11992-20) into the Luer-Lok fitting of one port in the manifold. Gently press a Spin Filter column into the PowerVac Mini Spin Filter Adapter until snugly in place. Ensure that all unused ports of the vacuum manifold are closed. Note: Aluminum PowerVac Mini Spin Filter Adapters are reusable if cleaned properly. 2. Transfer the 700 l of prepared sample lysate (from step 22 of the detailed protocol and step 6 from the short protocol) to the Spin Filter column. 3. Turn on the vacuum source and open the stopcock of the port. Hold the tube in place when opening the stopcock to keep the spin filter steady. Allow the lysate to pass through the Spin Filter column. Close the one-way Luer-Lok stopcock of that port. Note: If Spin Filter Columns are filtering slowly, close the ports to samples that have completed filtering to increase the pressure to the other columns. 4. Load 700 l of 100% ethanol into the Spin Filter so that it completely fills the column. Open the stopcock while holding the column steady. Allow the ethanol to pass through the column completely. Close the stopcock. 5. Add 600 l of Solution 4 to each Spin Filter. Open the Luer-Lok stopcock and apply a vacuum until Solution 4 has passed through the Spin Filter completely. Continue to pull a vacuum for another minute to dry the membrane. Close each port. 6. Turn off the vacuum source and open an unused port to vent the manifold. If all 20 ports are in use, break the vacuum at the source. Make certain that all vacuum pressure is released before performing the next step. It is important to turn off the vacuum at the source to prevent backflow into the columns. 7. Remove the Spin Filter column and place in the original labeled 2 ml Collection Tube. Place into the centrifuge and spin at 13,000 g for 1 minute to completely dry the membrane. 8. Transfer the Spin Filter column to a new 2 ml Collection Tube and add 50 l of Solution 5 to the center of the white filter membrane. Let stand at room temperature for 1 minute. Alternatively, sterile DNA-Free PCR Grade Water may be used for elution from the silica Spin Filter membrane at this step (MO BIO Catalog # 17000-10). 9. Centrifuge at room temperature for 30 seconds at 10,000 x g. 10
10. Discard the Spin Filter column. The DNA in the tube is now ready for any downstream application. No further steps are required. We recommend storing DNA frozen (-20 to -80 C). Solution 5 contains no EDTA. To concentrate the DNA see the Hints & Troubleshooting Guide. Thank you for choosing the UltraClean Standard Mini Plasmid Prep! 11
Hints and Troubleshooting Guide Concentrating the DNA Your final volume will be 50 l. If this is too dilute for your purposes, add 2 l of 5M NaCl and mix. Then add 100 l of 100% cold ethanol. Mix and incubate at -20 0 C for 1 hour. Centrifuge at 10,000 x g for 10 minutes. Decant all liquid. Dry residual ethanol in a speed vac or desiccator or ambient air. Resuspend the precipitated DNA in the desired volume. Amount of Culture to Process Use 2-5 ml of LB culture. To get higher yields use TB DRY Powder Growth Media (MO BIO Catalog# 12105). To Combine Cells from up to 5 ml of LB Culture Centrifuge 2 ml of culture for 30 seconds in a 2 ml tube. Discard supernatant. Repeat until 5 ml of culture has been processed into one tube. Low Recovery Two common causes of low yields: 1. The media was not completely removed. In this case, the pellet that results from spinning, after Solution 3 has been added, is not a firm white pellet on the side of the tube. The pellet is loose and it tends to be difficult to remove any liquid cell lysate. If you see this, start the prep over and be very careful to follow the recommended way to remove the culture media below. A good method to remove all the culture media: This method requires two spins. Be careful to orient the in the rotor the same way each time you spin them. Insert the in the centrifuge rotor so that the cap hinge is pointed straight out away from the center of the rotor. The first spin (1 minute) will pellet the cells. The large volume of culture media can then be dumped into a waste receptacle (typically a bottle containing bleach) by inverting the tube. The cap should then be replaced and the oriented in the rotor the same way. The second spin (5 seconds) will push all the residual media from the walls and cap to the bottom of the tube. If the tube was oriented the same as the original spin, the pellet will be very tight and the media will be easy to remove with a pipet tip. The best tips to use for this are the drawn out kind used to load sequencing gels. The small bore tip end makes removing very small volumes easier however; regular tips can also be used. The cells can now be resuspended in Solution 1 (see the notes below). 2. The cell pellet was not completely resuspended. In this case, a low yield results because clumps of cells do not lyse efficiently. There are a few good ways to resuspend the cells properly. A. Bump the tube on the vortex every 10 seconds during the 1 minute of vortexing it will take to resuspend the cells. Bumping means hold the tube on the vortex head for 10 seconds, take it off for 1 second then hold it on the vortex again. After 1 minute, hold the tube in a horizontal position up to a light and look at it. If you see any clumps of cells, keep bump vortexing until they are gone. It takes a minimum of 1 minute with the vortex at its highest speed to resuspend cells in the 50 l volume. Do two at a time when processing multiple preps. B. There is a paper in Biotechniques 1 that explains how cells can be resuspended in 5 seconds by scraping a tube across the holes of an 80 well microcentrifuge tube rack. The tube is held firmly in hand and the tube tip is run across the holes rapidly back and forth. It s easy and all you need is an 80 well rack. (They are great for storing, most labs have them already). 1. Voo, K. S.; Jacobsen, B. M. BioTechniques 24:240-243, February 1998 12
Hints and Troubleshooting Guide cont If DNA Floats Out of Well When Loaded on a Gel Residual Solution 4 in is in the final sample. Prevent this by being careful not to transfer Solution 4 liquid onto the bottom of the spin filter basket. Ethanol precipitate to remove residual Solution 4. See concentrating DNA above. Cleaning of the PowerVac Mini Spin Filter Adapters It is recommended to rinse the PowerVac Mini Spin Filter Adapters promptly after use to avoid salt build up. To clean the PowerVac Mini Spin Filter Adapters, rinse each adapter with DI water followed by 70% ethanol and flush into the manifold base. Alternatively, remove the adapters and wash in laboratory detergent and DI water. PowerVac Mini Spin Filter Adapters may be autoclaved. Do not use bleach to clean the PowerVac Mini Spin Filter Adapters while attached to the PowerVac Manifold. Bleach should never be mixed with solutions containing guanidine and should not be used to clean the PowerVac Manifold. For more information on cleaning the PowerVac Manifold, please refer to the PowerVac Manifold manual. 13
Contact Information Technical Support: Phone MO BIO Laboratories, Inc. Toll Free 800-606-6246, or 760-929-9911 Email: technical@mobio.com Fax: 760-929-0109 Mail: MO BIO Laboratories, Inc, 2746 Loker Ave West, Carlsbad, CA 92010 Ordering Information: Direct: Phone MO BIO Laboratories, Inc. Toll Free 800-606-6246, or 760-929-9911 Email: orders@mobio.com Fax: 760-929-0109 Mail: MO BIO Laboratories, Inc, 2746 Loker Ave West, Carlsbad, CA 92010 For the distributor nearest you, visit our web site at www.mobio.com/distributors 14
Other Quality Products Available from MO BIO Laboratories, Inc. For more product and detailed information go to www.mobio.com/catalog-request to request a catalog. DNA Purification and Gel Extraction Catalog No. Quantity RNA Isolation Continued Catalog No. Quantity PowerClean DNA Clean-Up 12877-50 RNA PowerSoil DNA Elution 12867-25 25 preps Accessory UltraClean 15 DNA Purification 12100-300 300 preps RNA PowerSoil Total RNA Isolation 12866-25 25 preps UltraClean PCR Clean-Up 12500-50 12500-100 UltraClean Microbial RNA Isolation 15800-50 15800-250 2 12500-250 2 UltraClean -htp 96 Well PCR Clean- 12596-4 4 x 96 preps UltraClean Tissue & Cells RNA 15000-50 Up UltraClean GelSpin DNA Extraction 12596-12 12400-50 12400-100 12400-250 12 x 96 preps 2 Isolation 15000-250 UltraClean Plant RNA Isolation 13300-20 13300-50 2 20 preps Plasmid DNA Isolation Catalog No. Quantity Genomic DNA Isolation Catalog No. Quantity UltraClean 6 Minute Mini Plasmid 12300-50 PowerLyzer PowerSoil DNA 12855-50 Prep 12300-100 Isolation UltraClean Standard Mini Plasmid Prep 12300-250 12301-50 12301-100 12301-250 UltraClean -htp 96 Well Plasmid Prep 12396-4 12396-12 UltraClean Midi Plasmid Prep 12700-20 12700-50 2 2 4 x 96 preps 12 x 96 preps 20 preps PowerLyzer UltraClean Microbial DNA Isolation 12255-50 PowerBiofilm DNA Isolation 24000-50 PowerFood Microbial DNA Isolation 21000-50 21000-100 UltraClean Maxi Plasmid Prep 12600-10 10 preps BiOstic Bacteremia DNA Isolation 12240-50 12600-20 20 preps UltraClean Endotoxin-Free Mini 12311-100 BiOstic FFPE Tissue DNA Isolation 12250-50 Plasmid Prep 12311-250 2 UltraClean Endotoxin-Free Midi 12711-10 10 preps BiOstic Paraffin Removal Reagent 12251-50 2 x 25 ml Plasmid Prep UltraClean Endotoxin-Free Maxi 12611-10 10 preps PowerMax Soil DNA Isolation 12988-10 10 preps Plasmid Prep UltraClean Endotoxin Removal 12615 1 kit PowerSoil DNA Isolation 12888-50 12888-100 UltraClean Endotoxin-Free Ethanol Precipitation 12616 1 kit PowerSoil -htp 96 Well Soil DNA Isolation 12955-4 12955-12 4 x 96 preps 12 x 96 preps UltraClean Endotoxin Removal Reagent 12625-25 25 ml UltraClean Soil DNA Isolation 12800-50 12800-100 Endotoxin-Free Sodium Chloride 12626-15 15 ml UltraClean -htp 96 Well Soil DNA Isolation 12896-4 12896-12 4 x 96 preps 12 x 96 preps Endotoxin-Free Centrifuge Tubes 12617-100 100 each/2 ml UltraClean Mega Soil DNA Isolation 12900-10 10 preps 12618-50 12619-25 50 each/15 ml 25 each/50 ml RNA Isolation Catalog No. Quantity PowerClean DNA Clean-Up 12877-50 PowerLyzer UltraClean Tissue & Cells RNA Isolation 15055-50 UltraClean Fecal DNA Isolation 12811-50 12811-100 PowerLyzer UltraClean Plant RNA 13355-50 PowerMicrobial Midi DNA Isolation 12225-25 25 preps Isolation PowerBiofilm RNA Isolation 25000-50 PowerMicrobial Maxi DNA Isolation 12226-25 25 preps LifeGuard Soil Stabilization Solution 12868-10 12868-100 12868-1000 12868-7500 On-Spin Column DNase I (RNase- Free) BiOstic Stabilized Blood RNA Isolation BiOstic Blood Total RNA Isolation 10 ml 100 ml 1 L 7.5 L UltraClean Microbial DNA Isolation 15100-50 UltraClean -htp 96 Well Microbial DNA Isolation 12231-20 20 preps 12231-50 12231-100 12230-20 12230-50 20 preps 12224-50 12224-250 10196-4 10196-12 PowerPlant DNA Isolation 13200-50 13200-100 UltraClean Plant DNA Isolation 13000-50 13000-250 2 4 x 96 preps 12 x 96 preps 2 15
Other Quality Products Available from MO BIO Laboratories, Inc. For more product and detailed information go to www.mobio.com/catalog-request to request a catalog. Genomic DNA Isolation Continued Catalog No. Quantity UltraClean -htp 96 Well Plant DNA 13096-4 4 x 96 preps Isolation 13096-12 12 x 96 preps UltraClean Tissue & Cells DNA Isolation UltraClean -htp 96 Well Tissue DNA Isolation UltraClean Blood DNA Isolation (Non-Spin) UltraClean Blood DNA Isolation (Processes 1,000 ml of Blood) UltraClean Blood DNA Isolation Plus RNase (Processes 1,000 ml of Blood) UltraClean BloodSpin DNA Isolation UltraClean -htp 96 Well BloodSpin DNA Isolation UltraClean Forensic DNA Isolation PowerWater DNA Isolation RapidWater DNA Isolation UltraClean Water DNA Isolation (0.45µm filters) UltraClean Water DNA Isolation (0.22 µm filters) UltraClean Water DNA Isolation (No filters) 12334-50 12334-250 12996-4 12996-12 2 4 x 96 preps 12 x 96 preps Other Reagents and Lab Accessories Catalog No. Quantity LB Agar (Lennox) Powder Growth 12109-05 Media, ph 7 12109-1 Soybean-Casein Digest Medium (TSB), USP Soybean-Casein Digest Agar Medium (TSA), USP 12109-5 12114-05 12114-1 12114-5 12115-05 12115-1 12115-5 12000-100 Yeast Extract 12110-05 12110-1 12110-5 12000-1000 1 kit Tryptone 12111-05 12111-1 12111-5 12002-1000 1 kit Agar, Bacteriological Grade 12112-05 12112-1 12112-5 12200-50 12200-250 12296-4 12296-12 14000-10 14000-20 14900-50-NF 14900-50-22 14900-50-45 14900-100-NF 14900-100-22 14900-100-45 14810-50-NF 14810-50-22 14810-50-45 14810-100-NF 14810-100-22 14810-100-45 14800-10 14800-25 14880-10 14880-25 14800-10-NF 14800-25-NF 2 4 x 96 preps 12 x 96 preps 10 isolations 20 isolations (No filters) (0.22 μm) (0.45 μm) (No filters) (0.22 μm) (0.45 μm) (No filters) (0.22 μm) (0.45 μm) (No filters) (0.22 μm) (0.45 μm) 10 preps 25 preps 10 preps 25 preps 10 preps 25 preps Microbiological Culture Media Catalog No. Quantity TB DRY Powder Growth Media 12105-05 12105-1 12105-5 LB Broth Powder Growth Media, ph 7 12106-05 12106-1 12106-5 LB Agar Powder Growth Media, ph 7 12107-05 12107-1 12107-5 LB Broth (Lennox) Powder Growth Media, ph 7 12108-05 12108-1 12108-5 20 bp DNA Ladder 17020-40 40 µg 100 bp DNA Ladder 17100-40 40 µg 1 kb DNA Ladder 17200-100 100 µg UltraClean Agarose, Molecular Biology Grade 15003-50 15003-100 15003-500 15003-1000 UltraClean MS-8 Agarose 15515-50 15515-100 15515-500 UltraClean Forensic Agarose 15505-50 15505-100 15505-500 UltraClean Low Melt Agarose 15005-50 15005-100 15005-500 UltraClean Low Melt Sieve Agarose 15004-50 15004-100 15004-500 Ethidium Bromide Solution 15006-1 15006-10 Ethidium Bromide Destaining Tea Bags Bromophenol Blue Gel Loading Buffer Bromophenol Blue/Xylene Cyanol Gel Loading Buffer 50 g 100 g 50 g 100 g 50 g 100 g 50 g 100 g 50 g 100 g 1 ml 10 ml 15007-25 25 bags 15008-1 15008-5 15009-1 15009-5 TAE Buffer, 50X (Tris-acetate-EDTA) 15001-100 15001-500 15001-1000 1 ml 5 x 1 ml 1 ml 5 x 1 ml 100 ml 500 ml 1 liter 16
Other Quality Products Available from MO BIO Laboratories, Inc. For more product and detailed information go to www.mobio.com/catalog-request to request a catalog. Other Reagents and Lab Accessories Continued Catalog No. Quantity Instrumentation and Accessories Catalog No. Quantity TBE Buffer, 10X (Tris-borate-EDTA) 15002-100 15002-500 15002-1000 100 ml 500 ml 1 liter PowerLyzer 24 Bench Top Bead- Based Homogenizer (110/220V) 13155 1 unit RNase-Free Gloves 1555-XS 1555-S 1555-M 1555-L UltraClean Lab Cleaner 12095-250 12095-500 12095-1000 KAPA PROBE FAST qpcr s 51220-100 51220-500 51220-1000 KAPA SYBR FAST Universal 2X 51230-100 qpcr Master Mix 51230-500 51230-1000 KAPA2G Robust HotStart ReadyMix 51240-100 51240-500 bag of 100 bag of 100 bag of 100 bag of 100 250 ml squeeze bottle 500 ml spray bottle 1 liter bottle 100 reactions 500 reactions 1000 reactions 100 reactions 500 reactions 1000 reactions 100 reactions 500 reactions PowerLyzer Tube Holder 13156 1 unit PowerLyzer Tube Holder Stand 13157 1 unit PowerVac Mini System 11992 1 unit + 20 adapters PowerVac Manifold 11991 1 unit PowerVac Mini Spin Filter Adapters 11992-10 11992-20 10 adapters 20 adapters KAPA HiFi HotStart ReadyMix 51250-100 100 reactions Ceramic Bead Tubes, 1.4 mm 13113-50 50 bead 51250-500 500 reactions KAPA2G FAST HotStart DNA 51260-100 100 reactions Ceramic Bead Tubes, 2.8 mm 13114-50 50 bead Polymerase with dntps 51260-250 51260-500 250 reactions 500 reactions KAPA2G FAST HotStart ReadyMix 51270-100 100 reactions Glass Bead Tubes, 0.5 mm 13116-50 50 bead 51270-500 500 reactions KAPA Long Range HotStart DNA 51280-100 100 reactions Glass Bead Tubes, 0.1 mm 13118-50 50 bead Polymerase with dntps 51280-250 51280-500 250 reactions 500 reactions KAPA Taq Polymerase ReadyMix 51290-250 250 reactions Metal Bead Tubes, 2.38 mm 13117-50 50 bead OmniTaq DNA Polymerase Enzyme 1224-250 250 reactions (10 U/µl) OmniTaq DNA Polymerase 2x 1226-250 250 reactions Master Mix (5 x 1.25 ml/tube) Omni KlenTaq DNA Polymerase 1225-250 250 reactions Enzyme (25 U/µl) Omni KlenTaq DNA Polymerase 2x 1227-250 250 reactions Master Mix (5 x 1.25 ml/tube) DNase (RNase-Free) 15600-5 5 mg 15601-100 2500 units Proteinase K 1223-100 100 mg 1222-2 2 ml (20 mg/ml) Ribonuclease A (25 mg/ml) 1202-1 1202-5 PCR Water 17000-1 17000-5 17000-10 17000-11 Molecular Biology Grade Water 17012-200 17012-5200 DEPC Treated Water 17011-200 17011-5200 Endotoxin-Free Water 17013-10 17013-50 17013-100 17013-500 1 ml 5 ml 1 ml 5 x 1 ml 10 x 1 ml 10 ml bottle 200 ml 5 x 200 ml 200 ml 5 x 200 ml 10 ml 50 ml 100 ml 500 ml 2.0 ml Tough Tubes with Cap 13119-500 13119-1000 500 1000 Carbide Bead Tubes, 0.25 mm 13121-50 50 x 0.5 ml Garnet Bead Tubes, 0.15 mm 13122-50 50 x 0.5 ml Garnet Bead Tubes, 0.70 mm 13123-50 50 x 2 ml Garnet + ¼ Ceramic 15 ml Bead Tubes, 0.70 mm Garnet + ¼ Ceramic 50 ml Bead Tubes, 0.70 mm ` 13134-50 50 13144-10 13144-50 13144-100 13144-500 10 50 100 500 Glass 15 ml Bead Tubes, 0.1 mm 13135-50 50 Glass 50 ml Bead Tubes, 0.1 mm 13145-10 13145-50 13145-100 13145-500 10 50 100 500 Glass 15 ml Bead Tubes, 1.0 mm 13136-50 50 Ceramic 15 ml Bead Tubes, 1.4 mm 13137-50 50 Ceramic 50 ml Bead Tubes, 1.4 mm 13147-10 13147-50 Metal 50 ml Bead Tubes, 2.38 mm 13149-10 13149-50 10 50 10 50 17
Other Quality Products Available from MO BIO Laboratories, Inc. For more product and detailed information go to www.mobio.com/catalog-request to request a catalog. Instrumentation and Accessories Continued Catalog No. Quantity Instrumentation and Accessories Continued Catalog No. Quantity PowerMix 15 ml Bead Tubes 13138-50 50 Whirl-Pak Collection Bag, Medium (1,627 ml) 23211-500 500 bags PowerMix 50 ml Bead Tubes 13148-10 13148-50 10 50 Whirl-Pak Collection Bag, Large (3,637 ml) 23212-250 250 bags 2 ml Collection Tubes 1200-100-T 1200-150-T 1200-250-T 100 150 250 2 ml Screw Cap Tubes 12800-200-E 200 & caps Whirl-Pak Stand up Bag, Small (118 ml) Whirl-Pak Stand up Bag, Medium (532 ml) 23220-500 500 bags 23221-500 500 bags 15 ml Collection Tubes 12700-T 25 Whirl-Pak Stand up Bag, Large (1,242 ml) 50 ml Centrifuge Tubes 12600-T 25 Whirl-Pak Stand up Bag, Extra- Large (2,041 ml) 23222-250 250 bags 23223-250 250 bags Spin Filters (in 1.9 ml ) 1200-50-SF 1200-100-SF 1200-250-SF Endotoxin-Free Centrifuge Tubes 12617-100 12618-50 12619-25 50 filters 100 filters 250 filters 100 each/2 ml 50 each/15 ml 25 each/50 ml Whirl-Pak Scoop Bag, 60 ml 23240-50 50 bags Anti-Static Funnels, Micro 23301-96 Pack of 96 15 ml Midi Spin Filters 12700-SF 25 spin filters Anti-Static Funnels, Small 23302-50 Pack of 50 Vortex-Genie 2 Vortex (120V) 13111-V 1 unit Anti-Static Funnels, Medium 23303-50 Pack of 50 Vortex-Genie 2 Vortex (220V) 13111-V-220 1 unit Anti-Static Funnels, Large 23304-20 Pack of 20 Vortex Adapter, holds 12 (1.5-2.0 ml) 13000-V1 1 unit Mini Horizontal Gel System 16001 1 each Vortex Adapter, holds 6 (5 ml) 13000-V1-5 1 unit Mini Horizontal Gel Caster, 3 place 16003 1 each Vortex Adapter, holds 4 (15 ml) 13000-V1-15 1 unit Mini Horizontal Gel Tray 16004 1 each Vortex Adapter, holds 2 (50 ml) 13000-V1-50 1 unit Polycarbonate Single-sided Comb 16005 16006 16007 16008 Vortex Adapter, holds 24 (1.5-2.0 ml) 13000-V1-24 1 unit Polycarbonate Dual-sided Comb 16013 16014 BagMixer 400 VW 23112 1 unit Teflon Single-sided Comb 16009 16010 16011 16012 BagFilter 400 P 23113-500 Box of 500 Teflon Dual-sided Comb 16017 BagPage 400 23114-500 Box of 500 Power Supply w/timer, (120V) 16023 1 unit 16015 16016 16018 16019 16020 1 mm x 3 well 1 mm x 8 well 1 mm x 10 well 1 mm x 12 well 1 mm x 8 well/16 well 1 mm x 10 well/14 well 2 mm x 8 well/16 well 2 mm x 10 well/14 well 1 mm x 3 well 1 mm x 8 well 1 mm x 10 well 1 mm x 12 well 1 mm x 8 well/16 well 1 mm x 10 well/14 well 2 mm x 8 well/16 well 2 mm x 10 well/14 well 18
Other Quality Products Available from MO BIO Laboratories, Inc. For more product and detailed information go to www.mobio.com/catalog-request to request a catalog. Instrumentation and Instrumentation and Accessories Continued Catalog No. Quantity Accessories Continued Catalog No. Quantity 96 Well Plate Shaker (120V) 11996 1 unit Vacuum Pump (120V) 11998 1 unit 96 Well Plate Shaker (220V) 11996-220 1 unit Vacuum Pump (220V) 11998-220 1 unit Plate Adapter Set 11999 1 set 19