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.
Bu some fans have speculated that Melisandre may have another, secret identity. According to one theory, she could be the child of a woman named Shiera Seastar, an illegitimate daughter of the Targaryen King Aegon IV, who lived many centuries before the era in which the current events of Game of Thrones are set.