酵母基因组DNA提取(Isolation of Yeast Genomic DNA)

 

　　 <center> <font>Amberg Lab ,Upstate Medical University DNA.html"> </font> <font> </font> <ol> <li> <font><font>Grow 100 ml culture to about 1x10 to the 8th </font> </font></li> <li> <font><font>Spin 4 x 15 mls of culture down about 3k x 3 min. </font> </font></li> <li> <font><font>Resuspend entire pool in about 6 mls ddH2O, aliqoute into 1.5ml microfuge tubes and spin down. </font> </font></li> <li> <font><font>Aspirate off super and vortex pellet till suspended. </font> </font></li> <li> <font><font>Add 200ul soln A, add 200ul Phenol/CHCl3 and 0.3g acid washed glass beads. </font> </font></li> <li> <font><font>Vortex 2 min, add 200ul TEpH8 and spin at max speed for 5 min. </font> </font></li> <li> <font><font>Transfer super to a new tube and add 1ml 100% EtOH, mix by inversion. </font> </font></li> <li> <font><font>Spin 2 min, pour off super and recon in 0.4ml TEpH8 30ug/ml RNaseA. </font> </font></li> <li> <font><font>Incubate 5 min x 37¡C, add 18ul 5M NH4OAc and 1ml 100% EtOH and store at -20¡C for a few hrs. </font> </font></li> <li> <font><font>Spin down DNA 10 min and recon each pellet in 25-50ul or whatever convenient volume. </font> </font></li> </ol> <ul> <li> <font><font>Solution A: </font> </font></li> <li> <font><font>200ul neat or 2ml 10% Triton X-100 (2%) </font> </font></li> <li> <font><font>1ml 10% SDS (1%) </font> </font></li> <li> <font><font>200ul 5M NaCl (0.1M) </font> </font></li> <li> <font><font>100ul 1M Tris pH8 (0.01M) </font> </font></li> <li> <font><font>20 ul 0.5M EDTA (1mM) </font> </font></li> <li> <font><font>Water to 10ml</font> </font></li> </ul> <div> <font>(责任编辑：admin)</font></div> </center>