DNA LADDR提取柱子如何选择

发布网友发布时间：2022-10-13 21:22

共1个回答

热心网友时间：2023-12-05 23:40

博凌科为解答：实验操作步骤：1、 Harvest cells and pre-cold PBS rinsing once, 500 g centrifuge 5 min Room Temprature2、 Pellet resuspended in 250 l TE buffer containing 1g/l RNase A3、 Cells were lysed by adding an equal volume of 1.2% SDS and gentle mixing by inversion(6-8 times)4、 Incubate 5 min, 350 l precipitation solution were added, mixed 8-10 times and put on ice for 15 min5、 Spin 14000g for 15 min Room Temperature and clear supernatant (700 l) transfer to miniprep column6、 Spin at 14000 g for 1 min, washed with 700 l wash buffer once7、 Spin empty column to discard the rest ethonal8、 30-50 l TE(pH8.0) elute the DNA fragments9、 1.6% agarose gel separated at 30V for 10min, and 80 V for 50 min10、 EB staining and photographed实验材料与试剂配制：1、 TE buffer2、 CsCl containing precipitation buffer:3M CsCl,1M Potassium acetate,0.67M acetic acid3、 Wash buffer:80mM potassium acetate,10mM Tris-Cl,pH7.5,40uM EDTA,60% ethanol4、 1.6% Agarose gel5、 Miniprep column (QIAprep Spin Miniprep Kit, QIAGEN)6、 ethidium bromide (10 l/100ml)我们实验室是这样做的，如果药物或者处理能够诱导凋亡，都还做得蛮好的，供参考。下面这个是VP16做的结果，还是很清楚的