Hello everybody
I am doing genotyping using KASP. My reaction show signs of contaminations in the NTC and across the samples. I am using new tip for every sample. Can somebody advice on how to detect DNA contamination? and how to remove it? and how to prevent it ?
Thank you
Hello Aisha,
Soumitesh already mentioned everything. The only thing I would add is to use a lab bench with uv radiation.
Best wishes
Dirk
Hi Aisha,
There are several ways to remove DNA contamination, which is a common nightmare for all PCR technicians. Here are some commonly used preventative measures, irrespective of whether your contamination is due to genomic DNA or carryover amplicons:
1. Make sure that your amplicon handling area and PCR set-up area are physically separated. It is ideal to have a DNA free environment for setting up PCR reactions i.e. a dedicated room where no one enters carrying any type of genomic DNA or amplicons
2. All the reagents used for PCR should be stored in that "clean" PCR room
3. Use filter-guard tips for minimization of aerosolic cross contamination of DNA
4. Use DNA-Zap to wipe off and decontaminate your PCR set up surface to get rid of any traces of amplicons or left over DNA
5. Change all the PCR reagents which resulted in contaminated products or replace one reagent at a time to identify which reagent carries the contaminant.
6. Use Uracil-DNA Glycosylase and dUTP for your PCR reactions to prevent any carry over amplicon contamination.
Hope this helps
Thank you so much for these advices. I think my problem is not taking care about the environment, I will follow your advice for removing DNA contamination.
Hello Aisha,
Soumitesh already mentioned everything. The only thing I would add is to use a lab bench with uv radiation.
Best wishes
Dirk
Thank you Dr Dirk
This is a good advice
I will follow all your advices.
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