In 1564 Sir James Melville, Ambassador of Mary, Queen of Scots (1542-87), was shown some portrait miniatures belonging to Elizabeth I. The English Queen 'took out the Queen's [Mary, Queen of Scots'] picture, and kissed it'. It is possible Melville was referring to this miniature and that it subsequently remained in the Royal Collection. The manner in which Elizabeth I treated the miniature indicates the intimate use of such small-scale paintings. By the reign of Charles I the miniature was mounted in a frame with eight other miniature portraits of James I's ancestors. This exquisite miniature is related to a drawing by François Clouet (c.1520-72) in the Bibliothèque nationale de France, Paris, but the drawing may be dated slightly earlier to c.1555. The clothing is updated in the miniature, in particular the detail of the ruff in the drawing which is changed into an open, standing collar. The drawing is only bust-length, whereas the miniature includes the sitter's hands, perhaps indicating their significance to the function of this portrait. The gesture of placing a ring on the fourth finger of the right hand is thought to allude to the marriage of Mary, Queen of Scots to the French Dauphin, the future Francis II, in 1558. This was standard symbolism for matrimony - for instance, on 13 August 1514, at the proxy marriage of Mary Tudor (Mary, Queen of Scots' great-aunt) to Louis XII of France, the Princess is recorded having placed a gold ring 'on the fourth finger of her right hand'. Mary, Queen of Scots had been betrothed to the Dauphin since the age of 5, and from that moment onwards was raised at the French court. This portrait would most likely have been commissioned by the French royal family from their court artist to commemorate the royal marriage. The jewel-like colours and contrasts of blue background and pink dress emphasise the Scots Queen's renowned beauty, with sparkling hazel eyes and auburn hair. In a double portrait of Mary as Queen of France with Francis II, in the Livres d'heures de Catherine de Médicis (Bibliothèque nationale de France, Paris), Mary is crowned and her hands are joined in prayer. However, the face, hair dressed with pearls and costume with standing collar in the manuscript illumination derive from the present work, indicating that Clouet's miniature was an important prototype for portraits of the Scottish and French Queen.