Related Manuals for Omega Mag-Bind M2837-00
Summary of Contents for Omega Mag-Bind M2837-00
- Page 1 Mag-Bind® Blood RNA 96 Kit 50 µL Blood M2837-00 1 x 96 preps M2837-01 4 x 96 preps 200 µL Blood M2839-00 1 x 96 preps M2839-01 4 x 96 preps May 2016...
-
Page 2: Table Of Contents
Mag-Bind® Blood RNA 96 Kit Table of Contents Introduction and Overview............2 Before Beginning................3 Quantification of RNA..............4 Kit Contents/Storage and Stability.........5 Preparing Reagents...............6 50 µL Mag-Bind® Blood RNA 96 Protocol (M2837)....7 200 µL Mag-Bind® Blood RNA 96 Protocol (M2839)..12 Troubleshooting Guide.............17 Ordering..................18 Manual Revision: May 2016 Innovations in nucleic acid isolation... -
Page 3: Introduction And Overview
Introduction and Overview The Mag-Bind® Blood RNA 96 Kit is designed for rapid and reliable isolation of total and viral RNA from mammalian whole blood. The Mag-Bind Bead technology provides high-quality RNA, which is suitable for direct use in most downstream applications, such as amplifications and enzymatic reactions. -
Page 4: Before Beginning
Before Beginning Important Notes Please take a few minutes to read this booklet in its entirety to become familiar with the procedures. Prepare all materials required before starting to minimize RNA degradation. • Whenever working with RNA, always wear gloves to minimize RNase contamination. Use only clean RNase-free disposable plastic pipette tips and plastic ware for the supplied reagents. -
Page 5: Quantification Of Rna
Quantification of RNA Quantification and Storage of RNA To determine the concentration and purity of RNA, measure absorbance at 260 nm and 280 nm with a spectrophotometer. One OD unit measured at 260 nm corresponds to 40 μg/mL RNA. DEPC Water is slightly acidic and can dramatically lower absorbance values. We suggest that you dilute the sample in a buffered solution (TE) for spectrophotometric analysis. -
Page 6: Kit Contents/Storage And Stability
Kit Contents Product M2837-00 M2837-01 Purifications (50 µL blood) 1 x 96 4 x 96 Mag-Bind® Particles CNR 530 µL 2.2 mL RBL Buffer 7 mL 30 mL Proteinase K Solution 600 µL 2.4 mL RXT Wash Buffer 22 mL 88 mL RNA Wash Buffer II 25 mL... -
Page 7: Preparing Reagents
Preparing Reagents Dilute RNA Wash Buffer II with 100% ethanol as follows and store at room temperature. 100% Ethanol to be Added M2837-00 100 mL M2837-01 400 mL 100% Ethanol to be Added M2839-00 200 mL M2839-01 280 mL per bottle Dilute RXT Wash Buffer with 100% ethanol as follows and store at room temperature. -
Page 8: Μl Mag-Bind® Blood Rna 96 Protocol (M2837)
M2837 Mag-Bind® Blood RNA 96 Protocol Mag-Bind® Blood RNA 96 Kit Protocol (M2837) - 50 µL blood The following protocol is designed for isolating total RNA from 50 μL fresh whole blood. For best RNA quality, always use blood that has not been frozen. Frozen blood can be used with this protocol, however, RNA quality could be compromised as the result of freeze- thaw process. - Page 9 M2837 Mag-Bind® Blood RNA 96 Protocol Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® Particles CNR. Remove the plate containing the Mag-Bind® Particles CNR from the magnetic separation device. Add 200 µL RXT Wash Buffer to each well. Pipet up and down 20 times or shake for 2 minutes to mix thoroughly.
- Page 10 M2837 Mag-Bind® Blood RNA 96 Protocol 16. Leave the tube on the magnetic separation device for 3 minutes to air dry the Mag- Bind® Particles CNR. Remove any residual liquid with a pipettor. 17. Prepare the Mag-Bind® DNase I digestion mix as detailed in the table below: Note: If total nucleic acid (DNA and RNA) is desired, skip Mag-Bind®...
- Page 11 M2837 Mag-Bind® Blood RNA 96 Protocol 21. Let sit at room temperature for 5 minutes. 22. Place the plate on a magnetic separation device to magnetize the Mag-Bind® Particles CNR. Let sit at room temperature until the Mag-Bind® Particles CNR are completely cleared from solution.
- Page 12 M2837 Mag-Bind® Blood RNA 96 Protocol 32. Transfer the cleared supernatant containing purified RNA into a new RNase-free microplate. 33. Store eluted RNA at -70°C. Note: Any combination of the following steps can be used to help increase RNA yield. •...
-
Page 13: 200 Μl Mag-Bind® Blood Rna 96 Protocol (M2839)
M2839 Mag-Bind® Blood RNA 96 Protocol Mag-Bind® Blood RNA 96 Kit Protocol (M2839) - 200 µL blood The following protocol is designed for isolating total RNA from 200 μL fresh whole blood. For best RNA quality, always use blood that has not been frozen. Frozen blood can be used with this protocol, however, RNA quality could be compromised as the result of freeze- thaw process. - Page 14 M2839 Mag-Bind® Blood RNA 96 Protocol Place the plate on a magnetic separation device to magnetize the Mag-Bind® Particles CNR. Let sit at room temperature until the Mag-Bind® Particles CNR are completely cleared from solution. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® Particles CNR.
- Page 15 M2839 Mag-Bind® Blood RNA 96 Protocol 15. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® Particles CNR. 16. Leave the tube on the magnetic separation device for 3 minutes to air dry the Mag- Bind® Particles CNR. Remove any residual liquid with a pipettor. 17.
- Page 16 M2839 Mag-Bind® Blood RNA 96 Protocol 20. Add 400 µL RNA Wash Buffer II. Pipet up and down 20 times or shake for 5 minutes to mix. 21. Let sit at room temperature for 5 minutes. 22. Place the plate on a magnetic separation device to magnetize the Mag-Bind® Particles CNR.
- Page 17 M2839 Mag-Bind® Blood RNA 96 Protocol 31. Place the plate on a magnetic separation device to magnetize the Mag-Bind® Particles CNR. Let sit at room temperature until the Mag-Bind® Particles CNR are completely cleared from solution. 32. Transfer the cleared supernatant containing purified RNA into a new RNase-free microplate.
-
Page 18: Troubleshooting Guide
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 1-800-832-8896. Problem Cause Solution Incomplete resuspension Resuspend the magnetic particles by of magnetic particles vortexing before use Make sure samples are stored properly RNA degraded during and that the samples are processed... -
Page 19: Ordering
RNA Wash Buffer II (20 mL) PDR046 HiBind®, E.Z.N.A.®, and MicroElute® are registered trademarks of Omega Bio-tek, Inc. Qiagen®, QIAvac® and Vacman® are all trademarks of their respective companies. PCR is a patented process of Hoffman-La Roche. Use of the PCR process requires a license.