The current sequencing method uses fluorescent labeled ddNTP. The sequencer only detects the sequences with fluorescence signals. Due to the lack of fluorescence, primers are generally invisible to the sequencer. For PCR products, in order to obtain the sequence of PCR primers, PCR products can be used for bidirectional detection or cloning into a vector, and sequencing with the primers fused to the vector（Note that the primers can't be too close to the insert fragment.）
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 is my gene sequence different from the standard 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?