MELT™ Total Nucleic Acid Isolation System: a new technology for hands-free tissue disruption, RNA preservation and total nucleic acid purification

Abstract

Ambion's multi-enzymatic liquefaction of tissue (MELT™) Total Nucleic Acid Isolation System allows the hands-free disruption of several solid tissues in parallel without tissue grinding or the need for a polytron. Up to 10 mg of fresh or frozen tissue can be lysed at room temperature in 5–15 min using a combination of powerful catabolic enzymes and a potent small-molecule RNase inhibitor. The procedure recovers high-quality total RNA with two- to threefold better yield and 40% less hands-on time than present methods. Moreover, MELT reagents irreversibly inactivate RNases, allowing the storage of liquefied tissue for more than a week at room temperature without an appreciable loss of RNA integrity. RT-PCR and microarray expression profiling studies of MELT RNA reveal an excellent correlation with RNA extracted using popular methods, including those based on single-reagent phenol solutions and glass-filter purification. A simple alteration in the protocol also allows the recovery of genomic DNA.