To make it clear what I'm talking about, I use a protocol like the following: 1  Take cells out of -80C and thaw on ice for 5 min. 2  Add 1 ul (~500 ng) plasmid DNA to 50 ul cells, mix gently with pipette tip. 3  Leave on ice for 30 min. 4  Put in 42C water bath for 45 sec.
Meliodas is the captain of the Seven Deadly Sins and is the Dragon's Sin of Wrath. He is the owner of the renowned Boar Hat, and the main protagonist of the series. His Sacred Treasure is the demon-splitting sword, Lostvayne. Accelerator vs Meliodas.
Eminent modern architect renowned as 'the man who built Brasilia'. Oscar Niemeyer, the architect who designed much of Brazil's futuristic capital, Brasilia, and Rio's Sambadrome. Photograph: Felipe Dana/AP There was a story told by the mayor of the town of NiterÃ³i, across the bay from Rio de Janeiro, that expresses perfectly the epic stature of the Brazilian architect Oscar Niemeyer, who has died aged 104.