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.
Melisandre is a red priestess of R'hllor and a shadowbinder, hailing from the city of Asshai in the Further East of Essos. She has joined the entourage of Stannis Baratheon, Lord of Dragonstone, believing him to be Azor Ahai reborn, a hero destined to defeat the Great Other.
Barbara Joan Barbra Streisand is an American singer, songwriter, actress, and filmmaker. In a career spanning six decades, she has achieved success in multiple fields of entertainment and has been recognized with two Academy Awards, ten Grammy Awards including the Grammy Lifetime Achievement Award and the Grammy Legend Award, five Emmy Awards including one Daytime Emmy, a Special Tony Award, an American Film Institute award, a Kennedy Center Honors prize, four Peabody Awards, the Presidential