An efficient method for large-scale isolation of plasmid DNAs by heat-alkali co-denaturation.
A method is presented for efficient and large-scale isolation of plasmid DNAs from bacterial cells. Based on the cooperativity of heat and alkali actions, the method provides DNA preparations with high quality and yield (about 2 micrograms of DNA/ml culture), which are completely digestable by restriction enzymes and have a high transformation efficiency. Furthermore, the DNA preparations are extremely stable, and even through 4-year storage at -20 degrees C, the electrophorogram and transformation efficiency remain as high as before. The factors affecting the stability of various DNA samples are discussed.
Sun
N E
NE
Department of Biochemistry, Nanjing University, People's Republic of China.
Shen
B H
BH
Zhou
J M
JM
Yuan
J
J
Xu
X X
XX
Zhu
D X
DX
Han
K K
KK
D000433:1-Propanol; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004492:Edetic Acid; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010957:Plasmids / Q000302:isolation & purification*
D004269:DNA, Bacterial; D000433:1-Propanol; D004492:Edetic Acid