博文
【reshipment】MINI PREP BY BOILING
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- Pellet 1.5 ml of an overnight culture at 12,000 rpm in Eppendorf centrifuge at RT for 3 min.
- Resuspend bacterial pellet in 350 µl of STET buffer.
- Add 25 µl of freshly prepared solution of lysozyme (10 mg/ml in 10 mM Tris-Cl, pH 8).
Mix by vortexing for 3 sec. - Place the tube in a boiling water bath for exactly 40 sec.
- Centrifuge the lysate at 12,000 g for 10 min at room temp.
- Remove the glob of bacterial debris with a sterile toothpick.
- Add to the supernatant 40 µl of 2.5 M NaOAc (pH 5.2) and 420 µl of isopropanol.
Mix by vortexing and store the tube for 5 min at room temp. - Centrifuge at 12,000 g for 5 min at 4°C.
- Remove the supernatant and wash with 1 ml 70% ethanol.
- Dry the pellet and resuspend in TE (pH 8) with RNase A (20 µg/ml).
https://blog.sciencenet.cn/blog-4083-210245.html
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