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Description:
With the GenElute Plant Genomic DNA Purification Kit, high quality genomic DNA can be purified from a variety of plant species (Table 1).
The kit contains all the reagents, columns, and tubes necessary to isolate genomic
DNA from up to 100 mg of fresh (Figure 1) or 20 mg of freeze-dried plant tissue. Plant
tissue is disrupted by grinding in liquid nitrogen, and DNA is released with
detergent and chaotrope. Proteins, polysaccharides, and cell debris are eliminated
with a 10 minute precipitation procedure followed by centrifugation through
a filtration column, included in the kit. The genomic DNA is purified further
by a silica bind-wash-elute procedure in microcentrifuge spin columns. Purified
DNA (Figure 2)
is ready for downstream applications such as PCR (Figure 3), Restriction Endonuclease Digestions, Cloning and Southern Blots. |
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Protocol for GenElute Plant
Genomic DNA Miniprep Kits
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Features & Benefits
| Fast |
Less than 40 minutes (Table 2) from tissue to purified genomic DNA, including disruption in liquid nitrogen. |
| Yield |
Typical DNA yields of up to 20 µg per prep. |
| RNA-free |
Genomic DNA without RNase A digestion (Figure 1)
and
(Figure 2). |
| Economical |
40% more purifications per kit than market leader. |
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GenElute Plant Genomic DNA Purification Kits
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Product Code
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Product Name
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Purifications
per kit
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Technical
Manual .pdf
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Short Protocol
.pdf
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G2N10
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GenElute Plant Genomic DNA Miniprep-10 prep
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10
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X
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X
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G2N70
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GenElute Plant Genomic DNA Miniprep-70 prep
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70
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X
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X
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| G2N350
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GenElute
Plant Genomic DNA Miniprep-350 prep
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350
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X
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X
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Kit Contents
- Lysis Solutions
- Precipitation Solution
- Binding Solution
- Wash Solution
- Elution Solution
- Filtration Columns
- DNA Binding Spin Columns
- Collection Tubes
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Table 1.
Typical yields of genomic DNA isolated from various plant species per 100 mg of starting leaf tissue.
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MATERIAL |
TYPICAL YIELD
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| Corn
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7.5 µg
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| Dianthus tissue culture |
3.3 µg
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| Pepper
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3.1 µg
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| Rice |
5.9 µg
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| Soybean |
5.7 µg
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| Tobacco
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5.2 µg
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| Tomato
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6.2 µg
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| Tomato (20
mg freeze dried leaf tissue) |
5.7 µg |
| Wheat
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11.5 µg
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Genomic DNA isolated from 100 mg fresh tomato leaves using various kits.
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Figure 1. Purified genomic DNA (0.4 µg/lane) was analyzed on a 0.8% agarose gel. Sigma, membrane-based; Supplier A, resin-based; Supplier M, solution based; Supplier Q, membrane-based. The Sigma prep shows the best yield of high molecular weight DNA, with less shearing (downward smear) than supplier Q. Markers are lambda DNA digested
with EcoRI / Hind III digest.
Note: RNA contamination present in DNA isolated using the kits from both Supplier A and M (smear below 0.6 kb).
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Genomic DNA isolated from various plant species isolated with GenElute
Plant Genomic DNA Miniprep Kit.
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Figure 2.
Purified genomic DNA (0.4 µg/lane) was analyzed on a 0.8% agarose gel.
Note: The DNA prepared from each species migrates as a tight band with little downward smearing, indicating that the majority is high molecular weight (> 21 kb).
No low molecular weight material is detected (< 0.6 kb), indicating that RNA has been eliminated without the use of RNase A digestion. Markers are lambda DNA Eco RI / Hind III digest. |
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PCR amplification of a 500 bp product isolated from genomic DNA.
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Figure 3. Genomic DNA from soybean leaves was purified using the GenElute Plant Genomic DNA Miniprep Kit. A 5 µl aliquot of eluate was used as template in a 20 µl total PCR reaction for 30 cycles. A 5 µl aliquot of each PCR reaction was resolved on a 1% precast agarose gel (Product Code 5722). The PCR marker (M) used (Product Code P9577) ranged from
50 bp to 2 kb. |
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Comparison of Preparation Time
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Table 2.The prep time required using the GenElute Plant Genomic DNA Miniprep Kit, compared to that of three kits from other suppliers.
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