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Description : The EZgeneTM Plant RNA Midiprep Kit provides a convenient and rapid method for isolating total RNA from a variety of plant samples. The kit introduces a homogenization column to efficiently remove cell debris and simultaneously homogenize the lysate. In combination with ezBind RNA spin column, high quality RNA can be extracted from as much as 500 mg tissue. Typical yields are shown in Table I. The EZgeneTM Plant RNA Kits are ideal for processing multiple plant samples in less than one hour. The need for organic extractions is eliminated, making total RNA isolation fast, safe, and reliable. Purified RNA has OD260/280 ratios of 1.8-2.0 and is suitable for the following applications. Table 1 Yields obtained with EZgeneTM Plant RNA Midi Kit Arabidopsis sp 150 μg Tobacco leaves 200 μg Mustard leaves 150 μg Maize 140 μg All components of the EZgeneTM Plant RNA Kit should be stored at 22-25 oC. Under these conditions, RNA has successfully been purified and used for RT-PCR after 24 months of storage. Under cool ambient conditions, a precipitate may form in the Buffer LY. In case of such an event, heat the bottle at 37 oC to dissolve. Store Buffer LY at room temperature. Each ezBind RNA column can bind approximately 100 μg RNA. Using greater than 500 mg plant tissue usually will not dramatically improve yields and sometimes has adverse effects. Problems Causes Suggestions Little or no RNA eluted RNA remains on the column Repeat elution. Pre-heat DEPC-treated ddH2O to 70 oC prior to elution. Incubate column for 10 minutes with water prior to centrifugation. Column is overloaded Reduce quantity of starting material. Clogged column Incomplete disruption or lysis of plant tissue. Completely disrupt sample in liquid nitrogen. Increase centrifugation time. Reduce amount of starting material Precipitated RNA will not dissolve. High nucleic acid and polysaccharide content. Reduce amount of starting material. Generally it is best to start with 50-100 mg at first. To avoid RNA degradation, do not increase incubation time for resuspension. Degraded RNA Source Freeze starting material quickly in liquid nitrogen and store at -70 oC without thawing. Follow protocol closely, and work quickly. Make sure that β-mercaptoethanol is added to Buffer LY. Use RB Buffer as dissolvent instead of DEPC-treated ddH2O RNase contamination Ensure not to introduce RNase during the procedure. Check buffers for RNase contamination. Problem in downstream applications Salt carry-over during elution Ensure RNA Wash Buffer has been diluted with 100% ethanol as indicated on bottle. Diluted RNA Wash Buffer must be stored at room temperature. Repeat wash with RNA Wash Buffer. DNA contamination Co-purification of DNA Digest with RNase-free DNase and inactivate at 75 oC for 5 minutes. Low Abs ratios RNA diluted in acidic buffer or water DEPC-treated ddH2O is acidic and can dramatically lower Abs260 values. Use TE Buffer (pH 8.0) to dilute RNA prior to spec analysis. Catalog # Product Name Preps Price $ R6611-01 Plant RNA kit 50 160.00 R6311-02 Plant RNA kit 250 750.00 R6612-01 Plant RNA midi kit 10 80.00 R6612-02 Plant RNA midi kit 20 160.00 R6614-01 Plant RNA maxi kit 10 120.00 R6614-02 Plant RNA maxi kit 25 270.00 R6615-01 Plant RNA mega kit 2 160.00 R6615-01 Plant RNA mega kit 10 700.00 R6815-01 96-well plant RNA kit 4x96 780.00 R6815-02 96-well plant RNA kit 20x96 3300.00 R6411-01 Blood RNA mini kit 50 150.00 R6411-02 Blood RNA mini kit 250 680.00 R6412-01 Blood RNA midi kit 10 80.00 R6412-02 Blood RNA midi kit 25 180.00 R6414-01 Blood RNA maxi kit 10 120.00 R6414-02 Blood RNA maxi kit 25 270.00 R6812-01 96-well blood RNA kit 4x96 780.00 R6812-02 96-well blood RNA kit 20x96 3500.00Introduction
Storage and Stability
Binding Capacity
Trouble Shooting Guide
Related EZgeneTM Products
- Model: R6614-00
- Manufactured by: Biomiga
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