Protocol for Re-using of Qiagen tips

Song-Kun Shyue shsu at HEART.MED.UTH.TMC.EDU
Wed Jan 3 12:36:46 EST 1996


Hi bio-net user:
        
        recently I try to resue the Qiagen column (it is too expensive) and
I got very good result! The way I used is followed you can try it. If anyone
have any other idea please share with me, Thanks.

1. For a tip 100, after eluted you DNA wash the column with 5 ml elution
buffer to get rid of trace DNA left.

2. Wash column with 4 ml equilibrium buffer, after 1 ml of the buffer folw
out stop the flow by wraping with parafilm and seal top with parafilm too. 

3. Kept tip up until next use.

        
        I try two cloumns one follow the protocol above the other was dry
after I washed with equil. buffer. the first one gave me about 130 ug DNA
from 10 ml 2xYT culture! 

        One critical step for high yield is to lyse your cells completely
when P2 solution added. If not complete clear after P2 added, add some more
P2 solution. Most problem with low yield comes from the incomplete lyse of
cells. In addition, make sure the cells have DNA in it. Run 5 ul of lysate
on gel after P3 (low voltage, about 50 V, for 4 min let DNA runs into gel,
then used high V. to save time) to see if there is DNA in your sample before
load into Tip to save your time and money.

        Good luck to everyone!




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