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-   -   How to prevent DNA cross-contamination in DNA extraction???? (http://www.molecularstation.com/forum/dna-extraction-forum/87956-how-prevent-dna-cross-contamination-dna-extraction.html)

trunick 06-26-2013 12:11 PM

How to prevent DNA cross-contamination in DNA extraction????
 
I perform DNA extraction on multiple samples simultaneously and opening microcentrifuge tubes contaminates my gloves with blood and I can't keep on discarding gloves after each sample, what should I do to wash DNA from the gloves between different samples? :wacko:

luisillo 06-26-2013 03:54 PM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Use diluted bleach. Just be careful not to let the bleach get to the inside of the tubes as it may spoil the sample.

trunick 06-27-2013 09:11 AM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Thank you very much, so I do I dilute bleach and wash my hands between samples. Do you think that vinegar, which is a substitute for bleach, can work with this?

trunick 06-27-2013 09:15 AM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Thank you very much, so I do I dilute bleach and wash my gloved hands between samples? Do you think that vinegar, which is a substitute for bleach, can work with this?
Can I dip a piece of cotton in the bleach and wipe the contaminated part of the glove with it between samples?
Sorry for the duplicate but the time for editing was expired

luisillo 06-27-2013 02:11 PM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Yes, you should soak some cotton and wipe your gloves with it (the contaminated part and its surroudings).

You can also use 70% ethanol to wipe the contamination of the gloves. Even though is not as strong to decontaminate as bleach, it has the advantage of better smell and that its volatile, which means it will air dry quickly. I work with blood samples too, and I haven't had problem using 70% ethanol.

trunick 06-28-2013 11:21 AM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Oh, you've put my mind to rest; I used 70% ethanol but then I read that it does not prevent DNA cross-contamination so I freaked out. I'm glad that I can continue using Ethanol because bleach is bad for the nervous system as well as the mucus membranes.

luisillo 06-28-2013 05:56 PM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
In fact, ethanol is NOT the best to prevent cross-contamination but it works fine to clean small spills (like small blood stains or drops). I believe is like this for 70% ethanol: the larger the contamination the lesser its effectiveness.

As I told you before, ethanol has worked fine for me. Still, if you are not completely sure you can use negative controls for DNA extraction, this is, using sterile water instead of sample (blood in your case), to confirm that there are not cross-contamination.

Good luck

trunick 06-29-2013 08:29 AM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
You mean that I should make one with all the steps but instead of my 300ul blood sample I can add water and see if it gives DNA?
I'll try that for sure
Thank you

luisillo 07-02-2013 02:19 PM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Quote:

Originally Posted by trunick (Post 447606)
You mean that I should make one with all the steps but instead of my 300ul blood sample I can add water and see if it gives DNA?

Yes, that is what will assure you that you are not suffering cross contamination.

Good luck. I bet your result will be alright.

trunick 07-03-2013 11:21 AM

Re: How to prevent DNA cross-contamination in DNA extraction????
 
Quote:

Originally Posted by luisillo (Post 447632)
Yes, that is what will assure you that you are not suffering cross contamination.

Good luck. I bet your result will be alright.

Thank you very much I will surely do that :-)


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