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Obtaining Reliable PCR from Seed DNA

Epicentre Biotechnologies Launches DNA Extraction Tool for a Variety of Seed Types

• Les Hoffman
• ,
• Shannon Krueger

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Seeds with High Polyphenolic Content

Figure 3. PCR of DNA extracted from cottonseed Click Image To Enlarge +

Figure 3. PCR of DNA extracted from cottonseed

Cottonseed is known to be problematic for DNA isolations because of its polyphenolic content. We tested 10 mg samples of chipped cottonseed extracted with QuickExtract Seed Solution as before. Seed DNA was amplified with the PlantAmp PCR System using primers for a single-copy transcription factor gene, DREB1 (Figure 3).

Faithful amplification was seen with both 1:10 and 1:100 dilutions of cottonseed extracts. In addition, we have obtained consistent and reliable results from oil sunflower seed in both end-point and real-time PCR.

DNA from Other Plant Tissues

Use of the QuickExtract Plant DNA Solution for extraction of PCR-ready DNA from plant tissues, especially those with high polyphenolic content, is highly recommended. This solution has been used successfully with a variety of plants, including Arabidopsis, barley, maize, emmer, pepper, rice, spelt, spinach, soybeans, and wheat.

Conclusions

The presence of storage carbohydrates and polyphenols can interfere with successful amplification of DNA prepared from seeds. The QuickExtract Seed DNA Extraction Solution facilitates the extraction of PCR-ready DNA from a wide variety of monocot and dicot seeds, even those containing polyphenols such as cottonseed and sunflower. The method is quick and effective, and avoids having to sprout seeds in order to use them for DNA analysis. The PlantAmp PCR System provides reliable PCR from seed DNA that contains PCR inhibitors.