Enhancing PCR amplification of DNA from recalcitrant plant specimens using a trehalose-based additive

Area of Science:

• Molecular Biology
• Plant Science
• Genetics

Background:

• Plant DNA extraction can be hindered by inhibitory compounds, complicating PCR amplification.
• Difficult plant specimens, such as herbarium material, often yield DNA that is resistant to standard PCR techniques.

Purpose of the Study:

• To evaluate the efficacy of a novel PCR additive reagent (TBT-PAR) in overcoming DNA inhibition.
• To improve DNA amplification success rates from challenging plant sources.

Main Methods:

• A PCR additive reagent (TBT-PAR) composed of trehalose, bovine serum albumin (BSA), and polysorbate-20 was formulated.
• PCR amplification was attempted on DNA extracted from various plant materials, including fresh, silica-dried, and herbarium specimens from four plant families.

Main Results:

• Standard PCR techniques failed for DNA from several plant species and types.
• The addition of TBT-PAR enabled successful PCR amplification of previously unamplifiable DNA samples.
• This included DNA from Achariaceae, Asteraceae, Lacistemataceae, and Samydaceae species.

Conclusions:

• TBT-PAR is an effective additive for routine PCR, enhancing DNA amplification from difficult plant specimens.
• This method significantly improves the success rate of PCR for DNA extracted from herbarium samples and other inhibitor-containing plant tissues.