CTAB 植物核酸萃取溶液
CTAB Extraction Buffer
CTAB 植物核酸萃取溶液可以廣泛用於植物組織的 DNA 萃取，多糖類 (Polysaccharides) 以及多酚 (polyphenols) 通常是萃取植物 DNA 時最棘手的問題，CTAB 植物核酸萃取溶液利用其陽離子試劑 (cationic detergent CTAB, hexadecyltrimethylammonium bromide/ cetyltrimethylammounium bromide) 來去除多糖類和多酚並透過 Polyvinylpyrrolidone 來結合多酚，來達成降低萃取汙染物的步驟。
傳統的 CTAB 核酸萃取會先將樣品加入 CTAB 溶液，而後進行樣品均質化作用，接著離心將雜質及多糖類沉澱下來，取上清液再進行 chloroform 萃取及酒精沉澱等步驟即可完成萃取，使用 CTAB 來進行植物樣品萃取通常可以獲得非常純淨的產物。
CTAB 萃取植物 DNA 的建議實驗步驟
- Pulverize 100 mg of plant sample using a liquid nitrogen chilled mortar and pestle. Once processed, mix 100 mg of frozen powdered sample with 500 µl of CTAB Plant Extraction Buffer.
- Place the homogenate into a 60°C bath for 30 min.
- Centrifuge the homogenate for 10 minutes at 10,000 x g.
- Transfer the supernatant into a clean tube and add 5 µl of RNase (10 mg/ml in water) to the lysate. Incubate at room temperature for 15 minutes.
- Centrifuge for 5 minutes at 10,000 x g.
- Extract the lysate with equal volume of chloroform: isoamyl alcohol (24:1). Vortex for 5 seconds then centrifuge for a minute at 10,000 x g to separate the phases.
- Transfer the upper phase to a clean tube.
- Repeat step 7 until upper layer is clear. Transfer upper phase to a new tube.
- Add 0.7 volumes of isopropanol. Mix and incubate at -20°C for 15 minutes.
- Centrifuge for 10 minutes at 10,000 x g. Decant and wash the pellet with 70% ethanol.
- Decant without disturbing the pellet. Dry the pellet briefly in the SpeedVac (3 minutes at medium heat). Do not over dry the DNA.
- Resuspend the DNA in 50 µl of TE buffer.
如果希望不使用 chloroform 的方法，可以參考不需 chloroform 的植物專用萃取試劑套組 SynergyTMM Plant DNA Extraction Kit.
產品列表
| 貨號 | 產品 | 數量 | |
|---|---|---|---|
| CEB 125-01 | CTAB Extraction Buffer | 125 ml | |
| CEB 500-02 | CTAB Extraction Buffer | 500 ml |
CTAB Extraction Buffer
- 可有效去除 polysaccharides 和 polyphenols
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