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

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

sanger sequencing pcr long-range
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updated by trausch trausch • written by DKA • DKA

2 answers

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.

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written by swbarnes2 swbarnes2

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

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written by DKA • DKA

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

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written by swbarnes2 swbarnes2

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

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written by DKA • DKA

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

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written by trausch trausch

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