Undoubtedly these two ways are different. PCR product contains all the PCR fragments. there is a number of amplicon have variations in PCR product, for instance, incomplete amplification, which adds to the level of background noise in sequence and leads to a low confidence base call. Some sites in several fragments are mutate, however, these sites in most amplified fragments are correct. While PCR clone product is one PCR fragment linked into the vector. Thus, direct sequencing can not reflect the wrong pairing sites, but coloned product reflects the mispairing in indirect sequencing.
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?