Hi

I wish to isolate eggs from sheep faeces and subsequently extract DNA and run a PCR from them. Part of the egg isolation technique involves floating them on a dense liquid. The three commonly used floatation liquids are Zinc sulfate, saturated sugar and saturated salt solutions. I was wondering what are the effects of these substances on the performance of the PCR reactions ?

Depends on the concentration, though most DNA extraction methods yield relatively pure DNA. On the other hand PCR is relatively robust and if detection limit is not too much of an issue even relatively dirty samples yield products.

I assume that you mean parasite eggs? If you need it pure you can use cleanup methods after the floatation and subsequent disruption.

You can always rinse the stuff off the eggs, no? Rinse and centrifuge? I don't think any of those you listed will affect the DNA, and the other parts you need to remove anyways.

On 6/10/2010 at 4:58 PM, bren2010 said:

Hi

I wish to isolate eggs from sheep faeces and subsequently extract DNA and run a PCR from them. Part of the egg isolation technique involves floating them on a dense liquid. The three commonly used floatation liquids are Zinc sulfate, saturated sugar and saturated salt solutions. I was wondering what are the effects of these substances on the performance of the PCR reactions ?

I assume this is a human sheep or is it an actual sheep. lmfao