RESUMO
This rapid, equipment-free DNA isolation procedure using chromatography paper is a simple method that can be performed in less than 30 min and requires no wet lab experience. With minimal expense, it offers an affordable alternative for anyone wanting to explore biodiversity. It also provides an excellent option for use in classrooms or other activities that are time limited. The method works best for plants or lichens, producing stable DNA on Whatman® chromatography paper at room temperature, which can be eluted as needed.
Assuntos
Código de Barras de DNA Taxonômico , Código de Barras de DNA Taxonômico/métodos , DNA/isolamento & purificação , DNA/genética , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Plantas/genética , Cromatografia/métodos , Líquens/genéticaRESUMO
Chelex-based DNA extractions are well suited for student DNA barcoding research because they are simple, safe, and inexpensive and can be performed without specialized laboratory equipment, allowing them to be performed in classrooms or at home. Extracted DNA is stable in Chelex solution for at least a week at ambient temperature, allowing collection of DNA samples from remote students. These extractions provide quality DNA for many taxa and are optimal for barcoding invertebrates, especially in combination with novel cytochrome c oxidase I (COI) primer cocktails and PCR cycling conditions.
Assuntos
Código de Barras de DNA Taxonômico , Complexo IV da Cadeia de Transporte de Elétrons , Reação em Cadeia da Polimerase , Código de Barras de DNA Taxonômico/métodos , Animais , Complexo IV da Cadeia de Transporte de Elétrons/genética , Reação em Cadeia da Polimerase/métodos , Invertebrados/genética , Invertebrados/classificação , DNA/genética , DNA/isolamento & purificaçãoRESUMO
A common way to avoid predators is by use of camouflage, a strategy which the stick and leaf insects (Phasmatodea) have refined by appearing as leaves, sticks, lichen, and moss. Stick and leaf insects have perfected their camouflage by sequestering diet-based carotenoids within their exoskeleton. Visual and chemical details of such camouflage have likely been influenced through the millennia of co-evolution between these insects and the plants they mimic. It is this evolutionary struggle that has resulted in a plethora of morphological and chemical adaptations across the stick and leaf insect family tree. In this chapter we discuss prior stick and leaf insect carotenoid studies, proper rearing of specimens, and describe methods for preparation of insect exoskeleton and plant samples, carotenoid extraction and analysis.