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DNA Sequencing
Genetic Diagnostic
Secugen - Technical Solutions - Stuttering After Mononucleotide Stretches
Sequence Appearance.
Sequencing data quality is poor after stretches of 10 or more nucleotides of the same base due to mixed bases.
Example.
Stuttering
Possibles Causes.
Polymerase slippage during DNA synthesis. This is a recognized limitation of the Sanger method.
Treatment.
Sequence from the reverse direction.
Use a poly-mononucleotide primer with a degenerate base (wobble) at the 3’ end.