Preparation of Plasmid DNA : Mini-prep.

��. Introduction
Plasmid? Bacterial chormosome? ????? ???? ?? ??? DNA?? ??? ??? ????. Plamid? ? ?? ???? ?? ???, ??? baterial cell growth? ???? ?? ???? ??? ??? ?? ???? ??? ??? ??. Plsmid? ??? cloning vector? ????? bateria??? plamid? ????? ??? ?? ????. ????? ?????? ???? ??? alkaline? detergent? ???? ???.

��. Procedure
1.Incubate bacterial culture (5ml) overnight at 37��C with vigorous shaking
2.Take 1.5ml of culture into eppendorf (EP) tube.
3.Spin down with 6K for 5 min
4.Remove the medium
5.Add 100��l of alkaline lysis solution I & resuspend bacterial pellet with up & down.
6.Add 200��l of alkaline lysis solution II & mix well by inverting the tube.
7.Add 200��l of alkaline lysis solution III & mix well by inverting the tube.
8.Spin down with 13K for 10min.
9.Transfer the supernatant to new EP tube.
10.Add 400��l of phenol:CIAA(1:1) & vortexing.
11.Spin down with 13K for 3min.
12.Transfer upper phase to new EP tube
13.Add 2x volume of 100% EtOH
14.Spin down with 13K for 15min at 4��C
15.Remove supernatant
16.Wash the pellet with 70% EtOH & remove supernatant
17.Air-dry
18.Dissolve the pellet in 20��l of TE containing 20��g/ml of RNase A.
(5? ???? ?????)