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Related Manuals for Omega Mag-Bind FFPE RNA 96
Summary of Contents for Omega Mag-Bind FFPE RNA 96
- Page 1 Mag-Bind® FFPE RNA 96 Kit M2551-00 1 x 96 preps M2551-01 4 x 96 preps March 2018...
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Page 3: Table Of Contents
Mag-Bind® FFPE RNA 96 Table of Contents Introduction..................2 Principle.....................2 Starting Materials................2 Kit Contents..................3 Preparing Reagents/Storage..........4-5 Mag-Bind® FFPE RNA 96 Protocol..........6 Mag-Bind® FFPE RNA 96 Protocol with Xylene....10 Troubleshooting Guide..............15 Manual Revision: March 2018... -
Page 4: Introduction
Introduction The Mag-Bind® FFPE RNA 96 kit provide a rapid and easy method for the isolation of total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Due to fixation and embedding procedures, nucleic acids in FFPE samples are heavily fragmented and modified by formaldehyde. -
Page 5: Kit Contents
Kit Contents Product Number M2551-00 M2551-01 Purifications 1 x 96 4 x 96 Mag-Bind® Particles SC 2.2 mL 8.4 mL RML Buffer 35 mL 140 mL MFB Buffer 20 mL 80 mL GFC Buffer 10 mL 40 mL RNA Wash Buffer II 25 mL 2 x 50 mL LPA Buffer... -
Page 6: Preparing Reagents/Storage
Preparing Reagents Please take a few minutes to read this manual thoroughly to become familiar with the protocol before beginning the procedure. Prepare all materials required before starting to minimize RNA degradation. Wear gloves/protective goggles and take great care when working with chemicals Dilute RNA Wash Buffer II with 100% ethanol and store at room temperature. - Page 7 Storage and Stability All of the Mag-Bind® FFPE RNA 96 Kit components are guaranteed for at least 12 months from the date of purchase when stored as follows. Mag-Bind® DNase I and DNase I Digestion Buffer should be stored at -20°C. Mag-Bind® Particles SC and LPA Buffer should be stored at 2-8°C for long-term use.
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Page 8: Mag-Bind® Ffpe Rna 96 Protocol
M2551 Mag-Bind® FFPE RNA 96 Kit Protocol Mag-Bind® FFPE RNA 96 Kit Protocol - 96-well plates Materials and Equipment to be Supplied by User: • Nuclease-free 1.2 mL round-well plates • Nuclease-free microplates • Centrifuge with swing-bucket rotor capable of 4,000 x g •... - Page 9 M2551 Mag-Bind® FFPE RNA 96 Kit Protocol Add 25 µL Proteinase K Solution (20 mg/mL). Incubate at 55°C for 15-30 minutes with occasional mixing. If necessary, extend the incubation to 1-3 hours or until the tissue is completely lysed. Incubate at 80°C for 15 minutes. Immediately centrifuge at 4,000 x g for 5 minutes.
- Page 10 M2551 Mag-Bind® FFPE RNA 96 Kit Protocol 16. Place the plate onto the magnet separation device to magnetize the Mag-Bind® Particles SC. Wait 3-5 minutes or until all the Mag-Bind® Particles SC are cleared from solution. 17. Aspirate and discard the cleared supernatant. Remove any liquid drops from each well.
- Page 11 M2551 Mag-Bind® FFPE RNA 96 Kit Protocol 27. Aspirate and discard the cleared supernatant. Remove any liquid drops from tube. 28. Repeat Step 24-27 for a second RNA Wash Buffer II wash step. 29. Air dry the Mag-Bind® Particles SC by leaving the plate on the magnetic separation device for 10 minutes.
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Page 12: Mag-Bind® Ffpe Rna 96 Protocol With Xylene
M2551 Mag-Bind® FFPE RNA 96 Kit Protocol with Xylene Mag-Bind® FFPE RNA 96 Kit Protocol with Xylene - 96-well plates Note: The following protocol uses xylene to remove paraffin from the FFPE sample. Use fume hood and take proper protection during xylene extraction. Materials and Equipment to be Supplied by User: •... - Page 13 Mag-Bind® FFPE RNA 96 Protocol with Xylene Centrifuge at 4,000 x g at room temperature for 5 minutes to pellet the tissue. Note: If the tissue does not form a tight pellet, centrifuge for an additional 3 minutes. Carefully remove and discard the xylene without disturbing the pellets. Add 1 mL 100% ethanol to each well.
- Page 14 Mag-Bind® FFPE RNA 96 Protocol with Xylene 18. Add 200 µL MFB Buffer, 20 µL Mag-Bind® Particles SC, and 430 µL 100% ethanol. Mix thoroughly by vortexing or pipetting up and down 10-20 times. Note: If the RNA content from sample is expected low or miRNA is the target, add 10 µL LPA Buffer.
- Page 15 Mag-Bind® FFPE RNA 96 Protocol with Xylene 27. Incubate at 37°C for 15 minutes. 28. Add 225 µL GFC Buffer. Mix thoroughly by vortexing for 20 seconds or pipetting up and down 10-20 times. Note: GFC Buffer must be diluted with 100% ethanol prior to use. Please see instructions on Page 4.
- Page 16 Mag-Bind® FFPE RNA 96 Protocol with Xylene 38. Add 30-50 µL DEPC Water. Resuspend the Mag-Bind® Particles SC thoroughly by vortexing for 30 seconds or pipetting up and down 30 times. 39. Let sit at room temperature for 10 minutes. 40.
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Page 17: Troubleshooting Guide
Troubleshooting Guide Please use this guide to troubleshoot any problems that may arise. For further assistance, please contact our technical support staff, toll free, at 800-832-8896. Possible Problems and Suggestions Troubleshooting Guide Problem Likely Cause Suggestions Resuspend the magnetic particles by Incomplete re-suspension vortexing before use. - Page 18 Notes:...