My fig wasps samples were stored in 70% ethanol at 4℃ and utilized for genomic DNA extraction. The genes I used in PCR are 18S, 28S and COI. But some of the sequencing results are bacteria and fungi. Is there any method to avoid the impact of bacteria and fungi?
Thanks.
Hi Wang,
Bacterial/Fungal contamination might have been caused from
1.DNA extraction kit used by multiple lab members are primary suspect for contaminants.
2. Water used during final stage of extraction or during PCR reaction may be contaminated.
Methods to eliminate and/or reduce background impurities and noise includes
-UV irradiation,
-restriction endonuclease and
-treatment with ethidium monoazide (EMA).
Best wishes!
Dissecting the wasps and avoid using the digestive tract in the DNA extraction should reduce a lot the microbial DNA.
Also, you can use primers with higher phylogenetic specificity (e.g. animal-, arthropod- or insect-specific) instead of using universal or domain-wide primers.
Please check that the laboratory is contaminated and carry out the experiment under aseptic conditions
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