The Synergy™ Plant DNA Extraction Kit features a proprietary chemistry that combines a grinding/extraction matrix and buffer system that rapidly disrupts and removes contaminants from plant samples. The Kit is easy to use, requiring the user to simply add up to 50 mg plant tissue and 500 µl Plant Homogenization Buffer to 2 ml pre-filled Homogenization Tubes prior to bead beating. The homogenate is centrifuged to pellet cell debris and contaminants (such as polyphenols and protein), leaving the DNA in the supernatant. While this cleared lysate is suitable for many PCR applications, DNA can be further purified by alcohol precipitation, if necessary, for samples with very high levels of polyphenols. The entire process can take less than 45 minutes and involves a fraction of the steps required by other protocols. Synergy™ Plant DNA Extraction Kit generates greater yields than both commercial kits and traditional CTAB protocols, with equal or better purity, as measured by 260/280 and 260/230 ratios.
The Synergy™ Plant DNA Extraction Kit includes: 100 pre-filled Homogenization Tubes (each containing grinding resin and satellite), Plant Homogenization Buffer (60 ml), RNase A Solution (600 µl), and instructions.
The standard Synergy™ protocol is simple:
- Place up to 50 mg of tissue in Synergy™ pre-filled Homogenization Tube. Add 500 µl Plant Homogenization Buffer.
- Homogenize via bead beating (approximately 5K oscillations). Centrifuge to pellet contaminants and debris.
- Transfer the cleared lysate to a new tube and add 5 µl RNase Solution. Incubate for 5 minutes. At this point the lysate can be used for PCR. Otherwise, add 0.7 volumes of isopropanol and incubate at -20°C for 15 min.
- Pellet the DNA by centrifuging at 10,000g for 5 min. Wash twice with ice cold 70% ethanol. Briefly dry the DNA and resuspend in 20 µl TE buffer.