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DNA can easily aerosolize, contaminating reagents, plastic ware, and DNA samples. This contamination leads to problems with band(s) of expected size amplifying from samples they should not amplify from. For example a wild type band amplifying from a homozygous mutant. This is why including a no template control is so important to include, because it can detect DNA contamination and prevent you from make incorrect conclusions about the genotype of your mice.
If you are amplifying bands of expected sizes in the no template control and samples.
| Solution | Rationale |
|---|---|
| Use only filter pipette tips for DNA extractions and PCR | Prevents contamination of pipettes |
| Clean pipettes | Aerosolized DNA can contaminate plastic ware |
| Replace all PCR reagents | DNA contamination may be in any PCR reagent, so these need to be systematically replaced |
If you are amplifying no band in the no template control, but are in your samples, it is likely the DNAs are contaminated.
| Solution | Rationale |
|---|---|
| Use only filter pipette tips for DNA extractions and PCR | Prevents contamination of pipettes |
| Clean pipettes | Aerosolized DNA can contaminate plastic ware |
| Re-extract DNAs using new reagents | Once the contamination is cleared you can re-extract from new tissue samples |