MELTTM Total Nucleic Acid Isolation System: a new technology for hands-free tissue disruption, RNA preservation and total nucleic acid purification
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چکیده
Ambion’s multi-enzymatic liquefaction of tissue (MELTTM) 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 highquality total RNA with twoto 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.
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تاریخ انتشار 2005