Mag-Bind® E-Z Pure - Omega Bio-Tek
Contents
1. 16 17 18 19 Mag Bind E Z Pure 96 well Plate Protocol Let sit at room temperature for 2 3 minutes Place the plate on a magnetic separation device to magnetize the Mag Bind E Z Pure Let sit at room temperature until the Mag Bind E Z Pure is completely cleared from solution Transfer the cleared supernatant containing purified DNA to a new 96 well microplate and seal with non permeable sealing film Store the plate at 2 8 C if storage is only for a few days For long term storage samples should be kept at 20 C Mag Bind E Z Pure 384 well Plate Protocol Mag Bind E Z Pure 384 well Plate Protocol Materials and Equipment to be Supplied by User e 384 well PCR plate containing PCR samples up to 100 uL well Magnetic separation device for 384 well PCR plates Vortexer Multichannel pipettor Reservoirs e Sealing film 70 ethanol Elution Buffer Cat PDR048 or 10 mM Tris pH 8 5 TE Buffer 0 1 mM EDTA or diH20 Skirted 384 well PCR plate e Optional Oven capable of 37 C 1 Read the manufacturer s instruction manual for the magnetic separation device if provided 2 Place the 384 well PCR plate on the bench and measure the volume of the PCR reaction Transfer the sample to a skirted 384 well PCR plate 3 Shake the Mag Bind E Z Pure to resuspend any Mag Bind E Z Pure particles that may have settled Add 1 8 volumes Mag Bind E Z Pure to each well PCR Reaction Volum2. Place the plate on a magnetic separation device to magnetize the Mag Bind E Z Pure Let sit at room temperature until the Mag Bind E Z Pure is completely cleared from solution Transfer the cleared supernatant containing purified DNA to a new 384 well microplate and seal with non permeable sealing film Store the plate at 2 8 C if storage is only for a few days For long term storage samples should be kept at 20 C Troubleshooting Guide Please use this guide to troubleshoot any problems that may arise For further assistance please contact the technical support staff toll free at 800 832 8896 Possible Problems and Suggestions F Increase the number amplification cycles Low PCR product yield for PCR Smaller PCR product size Small PCR fragments normally give lower yield Ethanol residue During the drying step remove any liquid from bottom of the well Low yield Particle loss during the Increase magnetization time procedure Aspirate slowly DNA remains bound to F Increase elution volume beads Incomplete resuspension of the beads during elution Vortex or pipet up and down to fully resuspend the beads Primer Insufficient wash of the Wash the beads one more time with 70 carryover particles ethanol Non specific The size of the non amplification specific amplification products were products are larger than not removed 100 bp 70 ethanol must be stored at room Salt carryover Problems in tempe
3. Ca OMEGA Innovations in nucleic acid isolation bio tek Product Manual Mag Bind E Z Pure M1380 00 5mL M1380 01 50 mL M1380 02 500 mL January 2013 For research use only Not intended for diagnostic testing Mag Bind E Z Pure Table of Contents Introduction and OvervieW ssesssssssseeesessssssssseeesesssssssss Illustrated Pr tOCOl ssissssssisserserssirissiriisrssirseirssisinrsnin Kit Contents and PreparatiOns ccccsccsssscsssssscseeeseeseesneenees Storage and Stability sserrssirsernenannananssa Mag Bind E Z Pure 96 well Plate Protocol ccseseeeee Mag Bind E Z Pure 384 well Plate Protocol Troubleshooting Guide ssessersersersesrssrreressrsrrssrssssssssssssss Ordering INfOrmMatiOn ssesssesssssessessensssssssssssssssssnssesssnesnsenesnses Manual Revision January 2013 N OMEGA bio tek Innovations in nucleic acid isolation Introduction and Principle Omega Bio tek s Mag Bind E Z Pure Kit allows rapid and reliable isolation of PCR products with high recovery rates The system combines Omega Bio tek s proprietary chemistries with the reversible nucleic acid binding properties of magnetic beads that selectively binds PCR amplicons 100 bp and larger and eliminate excess nucleotides primers and small non targeted amplification products such as primer dimers This kit is designed for both manual and fully automated purification of PCR samples Purified PCR fragmen
4. e pL Mag Bind E Z Pure uL C a a 4 Pipet up and down 5 10 times or vortex for 30 seconds 5 Let sit at room temperature for 1 minute 10 11 12 13 14 T5 16 Mag Bind E Z Pure 384 well Plate Protocol Place the plate on a magnetic separation device to magnetize the Mag Bind E Z Pure Let sit at room temperature until the Mag Bind E Z Pure is completely cleared from solution Aspirate and discard the cleared supernatant Do not disturb the Mag Bind E Z Pure Add 30 uL 70 ethanol to each well Let sit at room temperature for 1 minute It is not necessary to resuspend the Mag Bind E Z Pure Aspirate and discard the cleared supernatant Do not disturb the Mag Bind E Z Pure Repeat Steps 8 10 for a second 70 ethanol wash step Leave the plate on the magnetic separation device for 10 15 minutes to air dry the Mag Bind E Z Pure Remove any residue liquid with a pipettor Note It is important to dry the Mag Bind E Z Pure before elution Residual ethanol may interfere with downstream applications Optional Incubating the plate at 37 C can speed up the evaporation Remove the plate from magnetic separation device Add 30 pL Elution Buffer or 10 mM Tris pH 8 5 TE Buffer 0 1 mM EDTA or diH O to each well Pipet up and down 20 times or vortex for 30 seconds Let sit at room temperature for 2 3 minutes 17 18 19 10 Mag Bind E Z Pure 384 well Plate Protocol
5. rature downstream applications Ethanol carryover Non specific amplification products larger than 100 bp are not efficiently removed from PCR products Ensure the beads are completely dried before elution 11 12 Ordering Information The following components are available for purchase separately Call Toll Free at 1 800 832 8896 Ee OOOO f one 96 well Microplate 500 pL 25 pk HiBind E Z N A and MicroElute are registered trademarks of Omega Bio tek Inc PCR is a patented process of Hoffman La Roche Use of the PCR process requires a license
6. s Storage and Stability Mag Bind E Z Pure is guaranteed for at least 12 months from the date of purchase when stored at 2 8 C Mag Bind E Z Pure 96 well Plate Protocol Mag Bind E Z Pure 96 well Plate Protocol Materials and Equipment to be Supplied by User e 96 well PCR plate containing PCR samples up to 100 uL well Magnetic Separation Device Recommended Cat MSD 01 Vortexer e Multichannel pipettor Reservoirs e Sealing film 96 well microplate for elution 70 ethanol Elution Buffer Cat PDR048 or 10 mM Tris pH 8 5 TE Buffer 0 1 mM EDTA or diH O e 96 well processing plate Note the type of collection plate to be used depends on the type of magnetic separation stand used For OBI s MSD 01 a 500 uL conical bottom microplate is recommended Cat EZ9604 02 Optional Oven capable of 37 C 1 Read the manufacturer s instruction manual for the magnetic separation device if provided 2 Place the 96 well PCR plate on the bench and measure the volume of the PCR reaction Determine if transferring the sample to a processing plate is required If necessary transfer the PCR reactions to a 96 well microplate Note PCR Reactions gt 20 uL will need to be transferred to a processing plate MSD 01 is not compatible with PCR plates If processing in a PCR plate a magnet compatible with PCR plates must be used Recommended V amp P Scientific VP 771H 3 Shake the Mag Bind E Z Pure
7. to resuspend any Mag Bind E Z Pure particles that may have settled Add 1 8 volumes Mag Bind E Z Pure to each well PCR Reaction Volume pL Mag Bind E Z Pure pL 10 11 12 13 14 15 Mag Bind E Z Pure 96 well Plate Protocol Pipet up and down 5 10 times or vortex for 30 seconds Let sit at room temperature for 1 minute Place the plate on a magnetic separation device to magnetize the Mag Bind E Z Pure Let sit at room temperature until the Mag Bind E Z Pure is completely cleared from solution Aspirate and discard the cleared supernatant Do not disturb the Mag Bind E Z Pure Add 200 uL 70 ethanol to each well Let sit at room temperature for 1 minute It is not necessary to resuspend the Mag Bind E Z Pure Aspirate and discard the cleared supernatant Do not disturb the Mag Bind E Z Pure Repeat Steps 8 10 for a second 70 ethanol wash step Leave the plate on the magnetic separation device for 10 15 minutes to air dry the Mag Bind E Z Pure Remove any residue liquid with a pipettor Note It is important to dry the Mag Bind E Z Pure before elution Residual ethanol may interfere with downstream applications Optional Incubating the plate at 37 C can speed up the evaporation Remove the plate from magnetic separation device Add 30 40 ul Elution Buffer or 10 mM Tris pH 8 5 TE Buffer 0 1 mM EDTA or diH O to each well Pipet up and down 20 times or vortex for 30 seconds
8. ts can be used for microarrays automated fluorescent DNA sequencing restriction enzyme digestion and other applications The Mag Bind E Z Pure magnetic particles technology provides a better solution for nucleic acid purification compared to centrifugation and vacuum based technologies The product can be easily scaled up while providing very user friendly handling procedures If using Mag Bind E Z Pure for the first time please read this booklet to become familiar with the procedures PCR products are first mixed with Mag Bind E Z Pure PCR products then selectively bind to the Mag Bind E Z Pure particles With two rapid wash steps trace contaminants such as nucleotides primers and small non targeted amplification products are removed Pure DNA is eluted in Elution Buffer or water Purified DNA can be directly used in downstream applications without the need for further purification Illustrated Protocol Measure the PCR Reaction Add Mag Bind E Z Pure and Mix Magnetize and Remove Supernatant Wash Twice with 70 Ethanol Dry Elute DNA Kit Contents and Preparations Kit Contents Product Number M1380 00 M1380 01 M1380 02 Preparations 96 well format 5 mL 50 mL 500 mL 100 uL 27 preps 277 preps 2 777 preps PCR Reaction Volume M1380 00 M1380 01 M1380 02 384 well format 5 mL 50 mL 500 mL 555 preps 5 555 preps 55 555 preps 10 uL 277 preps 2 777 preps 27 777 preps 14 uL 198 preps 1 984 preps 19 841 prep
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