Hi every one.I have been trying to sequence some of the newly isolated strains of actinobacteria but every time I send them for sequencing the results comes out to be pseudomonas or alkaligens.now the problem is that there is no apparent contamination,morphologically they are confirmed actinobacteria, I have used two sets of primers the universal primers starting with 24f and 27f respectively.Is it possible that the gel purification might be compromised???or the Sequencing itself has some problems???any hint
Your primers might be contaminated. Check for any bands on a negative control.
There can be any number of possibilities about what you are describing. Also how you are analyzing the data. Since, the question do not give any specific information, I think it would not be possible to provide any specific solutions.
Contamination of primers or other chemicals used for PCR is the most probable reason.
- Badges
- Science method