Sequencing product purification by ethanol precipitation Sample Clauses

Sequencing product purification by ethanol precipitation. A solution was made by mixing 20 μl of the cycle sequencing product with 50 μl of 100% ethanol, 4 μl EDTA (125 mM) and 1μl sodium acetate (125 mM). The mix was vortexed and incubated at -20 oC for 30 minutes. The mix was centrifuged at 13,000 rpm for 25 minutes and the supernatant was discarded. The DNA was re-suspended in 200 μl of 70% ethanol, vortexed briefly and centrifuged again at 13,000 rpm for 20 minutes. The supernatant was removed and the DNA was dried up fully in the tube left unsealed at room temperature. DNA was re-suspended in 10 μl formamide and loaded into an optical ABI prism® plate. The new solution was denatured at 95 oC for 5 minutes on a thermal cycler and subsequently loaded on the automated sequencer ABI PRISM 3130-Avant Genetic Analyser (Applied Biosystems, Life Technologies).
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