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.
The underworld Goddess Melinoe was considered the daughter of Persephone and Hades, queen and king of the underworld. She was called Khthonia, Of the Lower World, a name also applied to Demeter. She was said to be half dark and half light (like Hel), from Her dark father Hades and Her mother who represented light.