Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment

Area of Science:

• Molecular Biology
• Biotechnology

Background:

• Standard PCR methods can be prone to feedback loops and contamination.
• Methylation-specific enzymes offer potential for targeted DNA manipulation.

Purpose of the Study:

• To introduce a novel PCR method, Déjà vu PCR, for unidirectional DNA processing.
• To develop a DNA decontamination technique using methylation-dependent enrichment.

Main Methods:

• Utilized six nucleotides in PCR reactions.
• Incorporated two methyl-specific restriction enzymes for targeted digestion.
• Developed a "DNA diode" system for unidirectional DNA flow.

Main Results:

• Achieved consecutive amplification with the addition of a 5th and 6th base.
• Demonstrated methylation-dependent mitochondrial DNA enrichment.
• Created an effective DNA decontamination technique generating ephemeral DNA.

Conclusions:

• Déjà vu PCR enables controlled, unidirectional DNA manipulation in laboratory settings.
• The method provides a novel approach for DNA decontamination and enrichment.