Shefigem2010: Plasmid mini prep
Plasmid Mini Prep
The protocol for plasmid mini prep was taken from the handbook provided by QIAGEN who provided the mini prep kit:
1.) Resuspend pelleted bacterial cells in 250μL Buffer P1 and transfer to a micro-centrifuge tube.
2.) Add 250μL Buffer P2 and mix thoroughly by inverting the tube 4-6 times.
3.) Add 350μL Buffer N3 and mix immediately and thoroughly by inverting the tube 4-6 times.
4.) Centrifuge for 10mins at 13,000 rpm in a table-top microcentrifuge.
5.) Apply the supernatant from step 4 to the spin column provided in the kit by decanting or pipetting.
6.) Centrifuge for 60s. Discard flow-through.
7.) Wash the spin column by adding 0.75ml Buffer PE and centrifuging for 60s.
8.) Discard the flow-through, and centrifuge for an additional 1min to remove residual wash buffer.
9.) Place the spin column in a clean1.5ml microcentrifuge tube. To elute the DNA, add 50μL distilled and autoclaved water to the centre of the spin column, let stand for 1min , and centrifuge for 1min.