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文献和实验Plasmid DNA Isolation from Bacteria
mg/ml proteinase K, mix , and incubate 1 hr at 37EC. 3.Add an equal volume of phenol/chloroform and mix well by inverting the tube until the phases are completely mixed. CAUTION: PHENOL CAUSES SEVERE BURNS, WEAR GLOVES GOGGLES, AND LAB COAT
. 11. Precipitate the plasmid DNA with PEG solution (30% polyethylene glycol, 1.6 M NaCl) by adding 0.4 ml and incubating 1hr-overnight on ice. This step discriminates very large plasmid DNA from small nucleic acid fragments as only the larger
High Efficency Yeast Plasmid Rescue
protocol. (NOTE: using up to 20 µl will increase yield of transformants) Colony Plasmid Rescue Protocol adapted by Julia Sidorova and Linda Breeden from Hoffman and Winston, Gene , 1987, Vol. 57: 267-272.
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