Detection of heterozygous exon deletion by Sanger and Long-Range PCR

Question

[DKA]

Hello, May I ask is it possible to detect a heterozygous exon deletion (50 bps) by Sanger sequencing using the quality scores of the region? Also can it be detected by Long-Range PCR, please? What is the best method to detect this deletion, please? Thank you

Answer 1

[swbarnes2]

I wouldn't try to call such things unless I could eyeball the traces, and confirm the boundaries of the indel. I would not rely only on per base quality scores.

[DKA]

Thank you. but if I know the boundaries of the deletion in the chromatogram. How can I spot a heterozygous exon deletion there, please?

[swbarnes2]

Have you actually looked at what a trace looks like for a heterozygous indel?

[DKA]

I took a look into the Sanger's chromatogram and it seemed normal. how does the abnormality look like, please?

Answer 2

[trausch]

There are tools that try to decompose heterozygous mutations in Sanger chromatogram traces (e.g., tracy decompose). The example on the web front end of tracy shows a chromatogram trace for a heterozygous deletion: www.gear-genomics.com/indigo