Hi, this is Dr. Harini, Manager with a genetic diagnostic lab. Recently we have done Sanger sequencing for PKD1 gene variant in proband and tested the fetus of the proband for the same. I need a clarification of the sequencing information as mentioned in the image attached. Can someone please clarify the same.
-
-
No T insertion variant carried by the proband was detected at the corresponding PKD1 locus in the fetal sample.Originally posted by harini.k View Post
The weak A peak observed in the reverse sequencing chromatogram represents a sequencing artifact (trailing peak) rather than a true heterozygous T>A variant, as it does not exhibit the characteristic peak pattern of a heterozygous substitution and is not supported by the forward sequencing results. Therefore, this signal should not be interpreted as a genuine nucleotide substitution.
To further confirm the result, the following approaches are recommended:- Repeat reverse sequencing using a new PCR amplification and a fresh sequencing reaction.
- Design a short PCR amplicon (≤300 bp) flanking the insertion site.
- Perform cloning-based sequencing, if permitted by legal and ethical regulations.
- Alternatively, conduct next-generation sequencing (NGS) followed by manual inspection of the BAM files.
-
The immune system’s power comes from its genetic diversity, allowing myriad threats to be neutralized through first recognizing foreign antigens. That diversity is also what makes the immune system so difficult to study. Recent advances in sequencing technology and computational biology, however, are giving researchers new tools to understand immune responses and immune-related diseases in greater detail.
This convergence of genetics, immunology, and computation...09-01-2026, 05:41 AM
Comment