DMSO use in PCR

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

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
Emmona87
Garter
Garter
Posts: 2
Joined: Tue Dec 14, 2010 3:48 pm

DMSO use in PCR

Post by Emmona87 » Tue Dec 14, 2010 3:52 pm

Hi all,

I am using DMSO in my PCRs and to minimise any contamination issues I have UV-treating my master mix components and making the master mix in a laminar cabinet before adding primers and DNA separately. I just wanted to check if it was safe to UV treat the DMSO prior to adding to the master mix?

Thanks

magicsiew
Coral
Coral
Posts: 145
Joined: Sun Jan 24, 2010 3:07 pm

Post by magicsiew » Tue Dec 14, 2010 3:59 pm

Why do u want to UV treat your master mix? I am not sure will it affect the master mix, since there are dNTP in your master mix. And did you put it on ice during the whole period while UV-treating?

Emmona87
Garter
Garter
Posts: 2
Joined: Tue Dec 14, 2010 3:48 pm

Post by Emmona87 » Tue Dec 14, 2010 4:06 pm

I am having problems with contamination so I am trying to prepare the DNA and master mix in separate areas to minimise this and UV treat all equipment and reagents that can be treated to prevent contamination. UV-treatment is only a couple of minutes in a DNA-free cabinet. The Taq is held in a container which keeps it cold but the other reagents are placed in the cabinet without ice.

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

Post by canalon » Tue Dec 14, 2010 4:57 pm

Rather than UV-treating my reagents (never heard of doing that), I would start with a fresh batch of reagents, clean my pipettes (inside and outside, take it apart and clean everything with mildly acidic solution) and start working with filter tips with 2 different areas for DNA and master mix (even better: a different set of pipettes in each area if possible). No idea on the effect of UV on DMSO, but there is no recommendation to store away from light, so I guess it is not sensitive.

Interestingly Taq is quite thermostable (I wonder why that surprise people, it is exactly why you use Taq in the first place!), but not so for your dNTPs. And I would not UV treat them either.
Patrick

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

User avatar
biohazard
King Cobra
King Cobra
Posts: 776
Joined: Sun Apr 08, 2007 6:45 pm

Re:

Post by biohazard » Wed Dec 15, 2010 7:05 am

canalon wrote:No idea on the effect of UV on DMSO, but there is no recommendation to store away from light, so I guess it is not sensitive.


Our lab always stores DMSO protected from light. From the Internet I found couple of references stating that DMSO is "somewhat sensitive to light" or so. So it is possible that UV light has some effect on DMSO, though I do not know how long exposure is required before something noticeable happens.

Like Canalon suggested, I would also go for two separate working areas, two sets of (clean) pipettes and filter tips.

Post Reply

Who is online

Users browsing this forum: No registered users and 3 guests