Highly efficient purification of total RNA by combining the stringency of either guanidinium thiocyanate or guanidinium hydrochloride with the speed and purity of silica-membrane column purification.
RNA was isolated from 20 mg freeze-dried budding leaves of A. thaliana using ISOLATE II RNA Plant Kit. The RNA was split into twelve replicates and diluted in a 10-fold series. Real-time reactions were performed using primers for a fragment of UBQ10 gene and SensiFAST SYBR No-ROX One-Step Kit. The results illustrate the quality of the RNA extracted through the reproducibility of the amplification.
RNA was isolated from 20 mg freeze-dried budding leaves of A. thaliana using ISOLATE II RNA Plant Kit. The extracted RNA was diluted in a 2-fold serial dilution (lanes 1-7) and PCR was performed using MyTaq One-Step RT-PCR Kit. The results illustrate the quality of the RNA obtained, as it can be used for very sensitive cDNA synthesis and PCR without further purification.
The ISOLATE II RNA Plant Kit provides a simple, efficient column-based method for the isolation of total RNA from a wide variety of plant materials, including leaves, bark, roots and fruits, without the need for hazardous reagents such as phenol.
By combining the stringency of guanidinium thiocyanate lysis with the speed and ease-of-use of silica-membrane purification, the ISOLATE II RNA Plant Kit provides a fast method for the purification of high-quality total RNA from most plant cells, including plant tissues, leaves, bark, roots and fruits.
Some plant tissues such as maize endosperm and the mycelia of filamentous fungi do not lyse well in guanidinium thiocyanate and can solidify, resulting in poor yields and so guanidinium hydrochloride lysis buffer is also provided as an alternative. Residual amounts of contaminating DNA are selectively removed using a column, however for ultrasensitive applications, remaining DNA can be removed using RNase-free DNase I that is supplied with the kit.
The ISOLATE II RNA Plant Kit has been designed to deliver optimal performance in RT-qPCR in conjunction with either the SensiFAST cDNA Synthesis Kit and SensiFAST Real-Time PCR Kits, or the SensiFAST One-Step Real-Time RT-PCR Kits. Additionally, the ISOLATE II RNA Plant Kit can be used to purify samples prior to RT-PCR amplification using the Tetro cDNA Synthesis Kit and any enzyme from the Meridian PCR portfolio, including MyTaq DNA Polymerase.
| Reagent | 10 Preps | 50 Preps | |
| ISOLATE II Filters | 10 | 50 | |
| ISOLATE II RNA Mini Columns | 10 | 50 | |
| Collection Tubes (2 mL) | 30 | 150 | |
| Collection Tubes (1.5 mL) | 10 | 50 | |
| Lysis Buffer RLY | 10 mL | 25 mL | |
| Lysis Buffer RLS | 10 mL | 25 mL | |
| Wash Buffer RW1 | 15 mL | 15 mL | |
| Wash Buffer RW2 | 6 mL | 12 mL | |
| Membrane Desalting Buffer MEM | 10 mL | 25 mL | |
| Reaction Buffer for DNase I RDN | 7 mL | 7 mL | |
| DNase I, RNase-free (lyophilized) | 1 vial | 1 vial | |
| RNase-free water | 13 mL | 13 mL | |
| Appearance | Colorless | ||
| Application | RT-qPCR, End-point RT-PCR, Northern, dot and slot blotting, Array analysis, Poly A+ RNA selection, RNA-Seq | ||
| Sample type | Plant cells and tissues or filamentous fungi | ||
| Preparation time | Approximately 30 minutes for 6 preps | ||
| RNA size distribution | > 200 nucleotides | ||
| Elution volume | 60 µL | ||
| Yield | Up to 200 µg of RNA | ||
| Storage | Store lyophilized DNase I (RNase-free) at 4°C and reconstituted at -20°C.
All other kit components at room temperature (18-25°C) |
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| Functionality | Nucleic acid recovery >90% | ||
| DNase, RNase-free | No RNase activity detectable | ||
Cat. No. Size
BIO-52076 10 Preps
BIO-52077 50 preps
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