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.
Even though Summer is alive, Ghost cannot sense him, presumably because Summer is beyond the Wall. Ghost seems to like Melisandre and Val, and Melisandre warns Jon to keep Ghost close. Much like Grey Wind at the Red Wedding, Ghost senses the impending treachery of the Night's Watch, and lashes out.