ISOLATE II Plant DNA Kit provides a simple, efficient column-based method for the isolation of genomic DNA from a wide variety of plant materials, without the
20 mg freeze-dried budding leaves of A. thaliana were homogenized in a mortar and pestle (lanes 1-3) and with a rotor stator homogenizer (lanes 4-6). Genomic DNA was isolated using ISOLATE II Plant DNA Kit and a 1.4 kb fragment of AOS gene was amplified from the isolated DNA. The results illustrating that the ISOLATE II Plant DNA Kit gives consistent results with different extraction techniques.
Genomic DNA was isolated from 20 mg freeze-dried budding leaves of A. thaliana using ISOLATE II Plant DNA Kit and a 2-fold serial dilution of the isolated DNA (200 ng to 6.25 ng, lanes 1-6 respectively) was used to amplify a 1.4 kb fragment of AOS gene. The results illustrating that the ISOLATE II Plant DNA Kit gives high yields of high quality DNA, ideal for all downstream applications.
By combining the stringency of CTAB lysis with the speed and ease-of-use of silica-membrane purification, ISOLATE II Plant DNA Kit provides a fast method for the purification of high-quality genomic DNA from most plant cells, particularly those rich in polysaccharides, including leaves, bark, roots and fruits as well as dung, animal-fecal, soil and compost samples.
Some plant tissues do not lyse well in CTAB and so an SDS-based lysis buffer is also provided as an alternative. Residual amounts of RNA remaining can be removed using RNase treatment during lysis with the RNase A supplied in the kit.
ISOLATE II Plant DNA Kit shows excellent recovery of plant DNA when different homogenization techniques are used. The silica membrane in the columns is optimized to improve DNA binding to give high-yields of high-quality DNA, even from difficult samples such as freeze-dried budding leaves. The extracted DNA is ready to use for subsequent downstream reactions such as PCR.
ISOLATE II Plant DNA Kit has been designed to deliver optimal performance in qPCR in tandem with SensiFAST Real-Time PCR Kits and in end-point PCR with MyTaq DNA Polymerase.
| Presentation | 10 Preps | 50 Preps | 250 Preps |
|---|---|---|---|
| ISOLATE II Filters | 10 | 50 | 250 |
| ISOLATE II Plant Column | 10 | 50 | 250 |
| Collection Tubes (2 mL) | 20 | 100 | 500 |
| Lysis Buffer PA1 | 5 mL | 25 mL | 125 mL |
| Lysis Buffer PA2 | 4 mL | 20 mL | 100 mL |
| Precipitation Buffer PL3 | 1 mL | 10 mL | 25 mL |
| Binding Buffer PB | 6 mL | 30 mL | 125 mL |
| Wash Buffer PAW1 | 6 mL | 30 mL | 125 mL |
| Wash Buffer PAW2 | 6 mL | 25 mL | 50 mL |
| Elution Buffer PG | 13 mL | 13 mL | 30 mL |
| RNase A (lyophilized) | 1.5 mg | 6 mg | 2 x 15 mg |
| Appearance | Colorless | ||
| Application | qPCR, End-point PCR, Southern, dot and slot blotting, Genotyping, Restriction digestion, Next generation sequencing, Bisulfite conversion/methylation analysis | ||
| Sample type | Plant cells and tissues or filamentous fungi | ||
| Preparation time | Approximately 30 minutes for 6 preps | ||
| Elution volume | 60 µL | ||
| Yield | Up to 50 µg of DNA | ||
| Storage | Store lyophilized RNase I at 4°C and reconstituted at -20°C.
All other kit components at room temperature (18-25°C) |
||
| Functionality | Nucleic acid recovery >90% | ||
Cat. No. Size
BIO-52068 10 preps
BIO-52069 50 preps
BIO-52070 250 preps
Katharina Schulte, Technical College Zweibruecken, Germany
Karlsruhe, Institute of Technology, Germany
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