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  • Isolation of DNA, RNA and protein from the starlet sea anemone Nematostella vectensis.

Isolation of DNA, RNA and protein from the starlet sea anemone Nematostella vectensis.

Nature protocols (2013-04-13)
Derek J Stefanik, Francis S Wolenski, Lauren E Friedman, Thomas D Gilmore, John R Finnerty
ABSTRACT

Among marine invertebrates, the starlet sea anemone Nematostella vectensis has emerged as an important laboratory model system. One advantage of working with this species relative to many other marine invertebrates is the ease of isolating relatively pure DNA, RNA and protein. Nematostella can be raised at high densities, under clean culture conditions, and it lacks integumentary or skeletal structures that can impede the recovery of DNA, RNA or protein. Here we describe methods used in our lab to isolate DNA, RNA and protein from Nematostella embryos, larvae and adults. The methods described here are less expensive than commercial kits and are more easily scalable to larger tissue amounts. Preparation of DNA can be completed in ∼7 h, RNA preparation in ∼1.5 h and protein preparation in ∼1 h.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, for molecular biology
Sigma-Aldrich
Pepstatin A, ≥100,000 U/mg