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.
Paprika is a main character in Blue's Clues. Paprika is Mr. Salt's and Mrs. Pepper's first baby, born in the season one finale. Unlike her parents, she does not speak with a French accent; Paprika's voice is small and squeaky. Many of Paprika's belongings are made from typical food items, such as her orange rattle and peanut crib.