Cheap DNA source for PCR control

[Mega]

Hi everyone,

 

a professor of mine told me he'd need Stapphyllococus Aureus and Streptococcus Epidermis DNA to make a PCR as a positive control...

 

Buying from DSMZ would cost some 380€ for 5 ug, so he asked if I had an idea where to get it from cheaply. He would just need 100 ng or so, to be able to run a PCR...

 

 

Obviously, I have no idea, it is biosafety level 2 and I'm not interested in working with potential pathogenic stuff...

He often helped me so now I'd like to help him too...

 

 

Does anyone have an idea where he could get it from?

 

Best,

Andreas

 

[Andreas Sturm]

The second one is not Streptocc. but also Stapphyllococcus Epidermitis...

 

 

 

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[Cathal Garvey]

If you're working in an academic lab licensed for BSL2 work, it's not
really that dangerous provided you know what you are doing. BSL 2 means
"Hazardous to immunosuppressed or pregnant people, or harmful to
agriculturally/culturally important species other than humans",
generally. For example, S.aureus is BSL2 as it's a common cause of
lethal hospital infections, but it lives on everyone, every day. To an
uninjured, healthy person working in a properly equipped lab, it's not
a serious hazard.

Better yet though, S.epidermis is harmless, I'm pretty sure it's BSL1.
Although a close relative of S.aureus, it rarely causes infections and
it lives on human skin (hence the name).

Both species are common contaminants of agar plates, though because one
is potentially hazardous (even if not very), I suggest asking around
for a characterised strain in the university. I'd be surprised if
someone didn't have a lab strain of one or the other species in a
freezer somewhere.

[Ashley Heath]

What type of PCR? If the amplicon is relatively small, say for qPCR, you could also have a synthetic oligonucleotide target made.

[Mega]

The synthetic template is a nice idea :)

I guess he would have known if we had a strain at university... but gonna tell him.

[Andreas Sturm]

He's gonna ask the lab keeper for Stapph. Epidermitis, which he may have. I suggested asking him and he said he hasn't done that yet... So your proposal helped already.

If anyone knew a cheap way to get also S.Aureus DNA (not the bug, because we only have BSL-1!), that would be nice...

On Mon, Jun 24, 2013 at 4:01 PM, Mega <masters...@gmail.com> wrote:

The synthetic template is a nice idea :)

I guess he would have known if we had a strain at university... but gonna tell him.

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[Avery louie]

we wanted to do a test PCR at bosslab, so we found some fungus growing on a plate, ground it up with a pestle in some drain cleaner, and added some master mix and universal barcoding primers.  If you are just looking for positive controls, some kind of harmless fungus/microbe +universal primers (very cheap) might be the way to go!

If you want to go with fungi, dakota might be able to comment on the primers we used.

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[Andreas Sturm]

I think my proff really wants to have Sapphylococcus Aureus proof of existance,

he's doing kInd of a project with their skin-microflora. And to have a positive control, he needs something that is S.Aur. for sure...

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