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.
Sondra Locke. Sondra Louise Anderson (nÃ©e Smith; born May 28, 1944), professionally known as Sondra Locke, is an American actress and film director. She made her film debut in 1968 in The Heart Is a Lonely Hunter, for which she was nominated for an Academy Award for Best Supporting Actress.