Cleaning DNA from test tube

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
samkh918
Garter
Garter
Posts: 20
Joined: Thu May 01, 2008 9:27 pm

Cleaning DNA from test tube

Post by samkh918 » Mon May 12, 2008 4:28 pm

Hello,

I have used 30ml KIMBLE test tubes for DNA MaxiPrep application. I still have traces of DNA on the sides of the tube and I have tried washing it with high pressure water or leaving water in it overnight for the DNA to redisolve but none have worked and I still see the traces on the sides.
Should I use a brush or is there other more efficient means? Is there any better solution than water to redisolve the DNA?

Thanks for your help

User avatar
MrMistery
Inland Taipan
Inland Taipan
Posts: 6832
Joined: Thu Mar 03, 2005 10:18 pm
Location: Romania(small and unimportant country)
Contact:

Post by MrMistery » Mon May 12, 2008 5:51 pm

trizol?
"As a biologist, I firmly believe that when you're dead, you're dead. Except for what you live behind in history. That's the only afterlife" - J. Craig Venter

User avatar
canalon
Inland Taipan
Inland Taipan
Posts: 3909
Joined: Thu Feb 03, 2005 2:46 pm
Location: Canada

Post by canalon » Tue May 13, 2008 2:08 am

Tris 10mM at pH7.5 might be a little better, DNA do not resuspend too well in acidic solution.

But what do you want resuspend the DNA or simply degrade it. In the latter case a 0,1M HCl solution should do the trick. It might not resuspend the DNA, but it will be depurinated and will not interfere with any other experiments. And brushing or using a dishwasher could also be really efficient (in fact some lab dishwasher have an Acid rinse in their cycle).
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

snowcapk
Garter
Garter
Posts: 37
Joined: Fri Oct 26, 2007 2:01 am

Post by snowcapk » Tue May 13, 2008 3:56 am

Since it's a maxiprep, I don't think the OP wants to degrade the DNA, but rather to collect it.

I have had this problem before. If it's a large construct like a BAC that you are purifying, *do not vortex* to resuspend. Try heating the solution at ~37C for 20 min. This should cause the DNA to resuspend in solution. You may still see a white pellet there, but it's likely that the majority of the DNA has resuspended at that point, so you might be able to give up on what remains.

Try measuring the concentration of DNA in the supernatant as it is. You may find that you have plenty of recovered DNA in solution.

samkh918
Garter
Garter
Posts: 20
Joined: Thu May 01, 2008 9:27 pm

Re: Cleaning DNA from test tube

Post by samkh918 » Tue May 13, 2008 4:48 am

Thank you for your feedbacks.

Yes, it's after the MaxiPrep, so I have already collected my DNA and just want to get rid of these residues on the sides of the tube so that they won't contaminate my future experiments.
I will try the brush and detergent and if that's not successful, I will use the 0.1 HCl.

Thanks again for your help.

Post Reply

Who is online

Users browsing this forum: No registered users and 8 guests