Plant Total RNA Isolation Kit Plus Total RNA Purificaiton Kit for Plant Rich in Polysaccharides and Polyphenols
Kit Description:
Cat.No.RE-05021/05022/05024
For purification of total RNA from general plant samples containing high polysaccharide and polyphenol components.
Quickly extract high-quality total RNA from plant samples with high content of polysaccharides and polyphenols.
RNase-Free Using DNA-Cleaning Column
Simple—all operations are completed at room temperature
Fast—operation can be completed in 30 minutes
Safe—no organic reagent used 
Specifications
50 Preps, 200 Preps
The kit uses the spin column and formula developed by Foregene, which can efficiently extract high-purity and high-quality total RNA from various plant tissues with high polysaccharides or polyphenols content. It provides the DNA-Cleaning column that can easily remove genomic DNA from the supernatant and tissue lysate. RNA-only column can effectively bind RNA. The kit can process a large number of samples at the same time.
The entire system does not contain RNase, so the purified RNA will not be degraded. Buffer PRW1 and Buffer PRW2 can ensure that the RNA obtained is not contaminated by protein, DNA, ions, and organic compounds.
Kit components
|
Buffer PSL1, Buffer PS, Buffer PSL2 |
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Buffer PRW1, Buffer PRW2 |
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RNase-Free ddH2O, DNA-Cleaning Column |
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RNA-Only Column |
Features&advantages
■ Operation at room temperature (15-25℃) throughout the whole process, without ice bath and low temperature centrifugation.
■ Complete kit RNase-Free, no need to worry about RNA degradation.
■ Particularly suitable for the purification of RNA from plant samples of polysaccharides and polyphenols.
■ DNA-Cleaning Column specifically binds to DNA, so that the kit can remove genomic DNA contamination without adding DNase.
■ High RNA yield: RNA-only Column and unique formula can efficiently purify RNA.
■ Fast speed: easy to operate and can be completed within 30 minutes.
■ Safety: no organic reagent is required.
■ High quality: The purified RNA fragments are of high purity, free of protein and other impurities, and can meet various downstream experimental applications.
Product parameters
■ Downstream applications: first-strand cDNA synthesis, RT-PCR, molecular cloning, Northern Blot, etc.
■ Sample: Fresh or frozen plant tissues of polysaccharides and polyphenols
■ Dosage: 50mg plant tissue
■ Maximum RNA binding capacity of purification column: 80 μg
■ Elution volume: 50-200 μl
Kit application
It is suitable for the extraction and purification of total RNA from fresh or frozen plant tissue samples (especially fresh plant leaf tissue) with high polysaccharide and polyphenol content.
Work flow
Diagram
Plant Total RNA Isolation Kit Plus processed 50mg of fresh leaves of polysaccharides and polyphenols, and 5% purified RNA was tested by electrophoresis.
1: Banana
2: Ginkgo
3: Cotton
4: Pomegranate
Storage and Shelf life
This kit can be stored for 24 months under dry conditions at room temperature (15-25℃); if it needs to be stored for a longer time, it can be stored in 2–8℃.
Buffer PSL1 can be placed at 4℃for 1 month after adding β-mercaptoethanol (it is recommended to add it at the same time of experiment).
The column plugged
After the column plugged, the RNA yield is reduced or even impossible to purify the RNA, and the obtained RNA mass is low.
Common cause analysis:
1. Sample breaks are not thorough.
Sample breakage does not completely cause DNA-CLEANING COLUMN to be blocked, while affecting RNA yield and quality. We recommend rapid grinding operation in a sufficient liquid nitrogen when you broke the samples, Try to crush the sample cell wall, cell membrane and other tissue. For plant samples of polyol polysaccharides, we recommend that you use Plant Total RNA ISOLATION KIT PLUS.
2. When suction the separated sample supernatant with DNA-Cleaning Column, the possible cell fragmented precipitate may be inhaled.
The cell fragmented sediments taken will cause the RNA-ONLY Column which will be blocked when the RNA adsorption operation is performed (see the step 6). We recommend you carefullly when suction this supernatant to avoid cell debris being sucked.
3. Sample initial amount is too much.
Excessive sample usage will result in incomplete sample fragmentation or incomplete cell lysis by Buffer PSL1, resulting in blockage of the purification column during purification. Plant Total RNA Isolation Kit Each single purified operating sample is 50 mg. For plant samples of polyol polysaccharides, we recommend that you try Plant Total RNA ISOLATION KIT PLUS.
4. The temperature of the centrifuge is too low.
The entire RNA isolation and purification process is carried out at room temperature (20-25°C), except that the sample tissue is broken by liquid nitrogen.The temperature of some cryogenic centrifuges is lower than 20℃, which may cause blockage of DNA-Cleaning Column and/or RNA-Only Column. If this happens, set the centrifuge temperature to 20-25℃, and make sure the lysis mixture and/or the ethanol-added supernatant was preheated to 37°C.
No RNA extracted or RNA yield is low
There are usually many factors that affect the recovery efficiency, such as: sample RNA content, operation method, the elution volume, etc.
Analysis of common causes as below:
1.An ice bath or low temperature (4°C) centrifugation was performed during the operation.
Suggestion: Operate at room temperature (15-25°C) in the whole process, do not do ice bath and low temperature centrifugation.
2.The RNA has been degraded due to improper preservation of the sample or long-term preservation of the sample.
Recommendation: Freshly collected samples should be quickly frozen in liquid nitrogen, and then stored at -80°C for a long time,avoid repeated freezing and thawing of samples; or immediately soak the samples in RNA stabilizer RNAlater solution (animal samples).
3.Insufficient sample fragmentation and lysis lead to blockage of the purification column.
Suggestion: When grinding the tissue, please ensure that the tissue is sufficiently ground, and quickly transfer it to the pre-prepared Buffer PSL1 (confirm that the correct proportion of β-ME has been added, see step 1 of the procedure).
4.The eluent was added incorrectly.
Suggestion: Make sure that RNase-Free ddH2O is dripped into the middle of the purification column membrane.
5.The correct volume of absolute ethanol was not added to Buffer PSL2 or Buffer PRW2.
Suggestion: Please follow the instructions, add the correct volume of absolute ethanol to Buffer PSL2 and Buffer PRW2 and mix well before the kit is used.
6.The amount of tissue sample is inappropriate.
Suggestion: Use 50 mg of tissue per 500 μl of Buffer PSL1. Using too much tissue will reduce the amount of RNA extracted and the purity of the resulting RNA will also be reduced. We strongly recommend that the initial sample dosage should not exceed 50 mg per RNA extraction operation.
7.Inappropriate elution volume or incomplete elution.
Suggestion: The eluent volume of the purification column is 50-200 μl; if the elution effect is not satisfactory, it is recommended to extend the time at room temperature after adding preheated RNase-Free ddH2O, such as 5-10min.
8.The purification column has ethanol residue after washing with BufferPRW2.
Suggestion: If the empty tube is centrifuged for 1 min and there is still ethanol remaining after washing in Buffer PRW2, you can increase the time of the empty tube centrifugation to 2 min, or place the purification column at room temperature for 5 min to fully remove the residual ethanol.
9.The kit was used incorrectly.
Suggestion: For plant samples of polyphenolic polysaccharides, using common kits such as Plant Total RNA Isolation Kit may not be able to obtain ideal RNA samples. We recommend you to use Plant Total RNA IsolationKit Plus, which is specially designed for polyphenolic polysaccharide plant samples. A kit specially developed for extracting RNA from polyphenol and polysaccharide plant samples.
OD260/OD280 value is low
RNA elution with ddH2O and used for spectrophotometer readings results in low OD260/OD280 values. We recommend using 10 mM Tris-HCl, pH 7.5 (rather than RNase-Free ddH2O to elute RNA) to obtain relatively correct OD260/OD280 values, see “RNA Concentration and Purification Assays” on page 19.
The purified RNA is degraded
The quality of purified RNA is related to factors such as sample preservation, RNase contamination, and manipulation.
Analysis of common causes:
1.Tissue samples were not stored in time after collection.
Recommendation: If the tissue samples are not used in time after collection, please store them in liquid nitrogen at low temperature immediately or transfer them to -80°C for long-term storage after quick freezing in liquid nitrogen, or immediately immerse the samples in RNA stabilizer RNAlater solution (animal samples ). For RNA extraction, try to use freshly collected tissue samples.
2.Repeated freezing and thawing of tissue samples.
Suggestion: When storing tissue samples, it is best to cut them into small pieces for preservation, and take out a part of them when using them to avoid the degradation of RNA caused by repeated freezing and thawing of the samples.
3.RNase is introduced in the operation room or not worn disposable gloves, masks, etc.
Suggestion: RNA extraction experiments are best performed in separate RNA operations, and the laboratory table should be cleaned before the experiment, and disposable gloves and masks should be worn during the experiment to avoid RNA degradation caused by the introduction of RNase to the greatest extent.
4.The reagent is contaminated by RNase during use.
Suggestion: Replace with a new series of plant total RNA extraction kits for related experiments.
5.The centrifuge tubes and pipette tips used for RNA manipulation are contaminated with RNase.
Suggestion: Make sure that the centrifuge tubes, pipette tips, pipettes, etc. used in RNA extraction are all RNase-Free.
Instruction Manuals:
Plant Total RNA Isolation Kit Plus Instruction Manual
