There are several reasons.
1. Inadequate template concentration.
2. Not a single-colony.
3. Multiple priming sites present.
4. Inadequate primer concentration(customer provide primer)
5. Incorrect PCR products.
Articles in this section
- Will the primers degrade at room temperature during transportation? Can we use dry ice for transporting to maintain stability?
- Why there is a difference between my sequencing data and the one from the references?
- Why the short PCR product is not suitable for direct sequencing?
- Why target gene products are seen in control sample with no template?
- Why my sequence results are reverse complement to the template DNA, while I demand a 5’ to 3’ orientation?
- Why is the TM value for primers of Genscript different from the TM value I obtained from other software?
- Why is the sequence of primers in the sequencing results different from the expected sequence?
- Why I got a sequencing result that was not I want?
- Why does the sequencing primer need to be site-specific on the DNA template?
- Why does my sequencing fail?