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Description:
The Extract-N-Amp Plant PCR Kits contain all the reagents necessary to rapidly extract
genomic DNA from plant leaves (Figure
1) and amplify targets of interest by PCR. A novel Extraction Solution
eliminates the need for conventional freezing of plant tissues with liquid nitrogen,
mechanical disruption, organic extraction, column purification, or precipitation of
DNA. The kit also includes a PCR reaction mix, especially formulated for amplification
directly from the extract. This formulation uses an antibody based Hot Start for specific amplification. The PCR master mix comes in two formulations: Extract-N-Amp PCR Reaction Mix and REDExtract-N-Amp Plant PCR
Kit. The REDExtract-N-Amp™ PCR mix is formulated with
REDTaq, which acts as a tracking
dye and allows for convenient direct loading of PCR reactions onto agarose gels for
analysis.
Genomic DNA is extracted from 0.5 to 0.7 cm plant leaf disks that have been cut
with a standard paper punch and simply incubated in Extraction Solution at 95 °C for
10 minutes. An equal volume of Dilution Solution is added to the extract to neutralize
inhibitory substances prior to PCR. A portion of the DNA extract is then added to a
PCR reaction containing primers and either the REDExtract-N-Amp or Extract-N-Amp PCR.
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Features & Benefits
| Fast |
Single-step extraction of plant genomic DNA for PCR in less than 15 minutes without need for freezing, mechanical disruption, organic extraction, column purification, or precipitation. |
| Versatile |
Extract and amplify target DNA sequences from a variety of plant species (Figure 1). |
| High Specificity |
Hot Start antibody for highly specific PCR amplification of genomic DNAs. |
| Convenient |
Kits include PCR ready mixes, optimally formulated for use with leaf extracts. RedTaq PCR Ready Mix requires no loading buffers or tracking dyes for gel analysis. |
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Extract-N-Amp Plant PCR Kits
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Product Code
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Product Name
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Number of Extractions
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Number of Amplifications
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Technical Manual .pdf
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Extract-N-Amp Plant PCR Kit
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100
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100
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Extract-N-Amp Plant PCR Kit
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100
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500
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Extract-N-Amp Plant PCR Kit
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1000
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1000
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Extract-N-Amp Plant PCR Kit
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1000
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5000
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REDExtract-N-Amp Plant PCR Kits
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Product Code
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Name
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Number of Extractions
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Number of Amplifications
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Technical Manual .pdf
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REDExtract-N-Amp Plant PCR Kit
(contains REDTaq)
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10
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10
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REDExtract-N-Amp Plant PCR Kit
(contains REDTaq)
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100
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100
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REDExtract-N-Amp Plant PCR Kit
(contains REDTaq)
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100
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500
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REDExtract-N-Amp Plant PCR Kit
(contains REDTaq)
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1000
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1000
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REDExtract-N-Amp Plant PCR Kit
(contains REDTaq)
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1000
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5000
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| Kit Contents |
Extract-N-Amp Kit
- Extraction Solution
- Dilution Solution
- Extract-N-Amp PCR Ready Mix
- 2 ml Tubes for Extraction (not included with the 1,000 Amplification kit)
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REDExtract-N-Amp Kit
- Extraction Solution
- Dilution Solution
- REDExtract-N-Amp PCR Ready Mix
- 2 ml Tubes for Extraction
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PCR analysis of genomic DNA extracted from 5 different plant species using Sigma's Extract-N-Amp Plant Kit.
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Figure 1.
Extract-N-Amp Plant PCR Kit used to isolate and amplify genomic DNA from various plant sources. Genomic DNA was extracted from 0.5 cm leaf disks that were cut using a standard paper punch. DNA was extracted using the Extract-N-Amp Plant PCR Kit in less than 15 minutes. All samples were then amplified using the specially formulated Hot Start PCR
mix. The products were generated from a 30-cycle duplex reaction containing primers
specific to plant chloroplast (upper band) and primers specific to Cannabis sativa DNA
(lower band). MW ladder is 100, 200, 400 and 800 bp. Data provided by Andy Hopwood,
Forensic Science Service, Birmingham, England.
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REDTaq allows for convenient loading onto agarose gels
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Figure 2.
Samples are loaded directly on an agarose gel after PCR amplification with RedTaq DNA Polymerase, obviating the need or addition of loading buffers and/or tracking dyes. The RedTaq dye approximately co-migrates with a 125 bp duplex. |
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Stability of Extract-N-Amp Plant Extracts
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Figure 3.
Stability of DNA in corn leaf extracts. Eight disks were punched from a corn leaf, and DNA was extracted according to the procedure in the Technical Bulletin for the Extract-N-Amp
Plant Kit. Two 4 5l aliquots from each were analyzed immediately by quantitative PCR
with SYBR™ Green detection on an ABI Prism 7700. DNA standards for
quantitative PCR were purified DNA prepared from corn leaf tissue with the GenElute
Plant Genomic DNA Kit
(Product Code G2N70). Half of the leaf extracts
were stored at 4 °C (recommended storage conditions) and the other half at 37 °C (accelerated storage). Quantitative PCR was repeated after 1 week, 3 weeks, 6 weeks
and 6 months from extracts at 37 °C. Results for storage at 37 °C are shown.
The average of 2 replicate PCR assays from each extract is plotted. Error bars
represent one standard deviation.
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