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- Main article: Matilda Middleton
Matilda "Tilly" Middleton (born September 7, 1540) was the daughter of Mr. Middleton, a physician at the Tower of London. She had an elder brother, William, and five younger siblings, Jack, Harry, Mary, Susannah, and Edward. In 1553, her father gave her a diary, which contained a letter from Anne Boleyn to her daughter, the Princess Elizabeth. Tilly waited patiently for several years for a chance to give Elizabeth the letter.
- Main article: Elizabeth I
Elizabeth (September 7, 1533 – 1603) was the daughter of Henry VIII and Anne Boleyn. She was also the sister Mary I and Edward VI. In 1544, she was suspected of participating in Wyatt's rebellion and sent to live in Tower of London by Mary. She was later moved to Woodstock in May, and to her home in Hertfordshire in October 1555. Upon Mary's death in 1558, Elizabeth became Queen of England. Her coronation was held on January 14, 1559.
Frances Lea was the niece of Master Lea, who worked at the Mint in the Tower of London. She moved to London from the country. Frances became good friends with Tilly, despite getting Tilly into trouble frequently. She was described as "forward," and enjoyed playing pranks and telling jokes. Frances was also used to getting what she wanted, which sometimes caused friction between her and Tilly. By 1558, she appeared to have a crush on Tilly's brother, William
- Main article: Mary I
Mary (1516 – November 17, 1558) was the Queen of England from 1553 to 1558. She was the daughter of Henry VIII and Catherine of Aragon. Her brother, King Edward VI attempted to disinherit Mary by naming their cousin, Lady Jane Grey as his successor. Nonetheless, Mary was crowned and had Jane executed in early 1554. Mary married a Catholic prince, Philip of Spain in July 1554. She had many Protestants burned at the stake for heresy throughout her reign. Mary never produced an heir, leaving the throne to Elizabeth upon her death.
Master Middleton was the father of William, Tilly, Jack, Harry, Susannah, Mary, and Edward. He worked as a physician at the Tower of London, and attended to Lady Jane Grey and Princess Elizabeth, among others. He disliked to speak of his work or patients while at home. Years earlier, he received a diary from Anne Boleyn, which he later gave to his daughter, Tilly. His eldest son, William, worked as his apprentice.
Mrs. Middleton was the mother of William, Tilly, Jack, and Harry. In 1553, she gave birth to a daughter, Susannah, who died shortly after. The following year, she gave birth to twins, Mary and Susannah "Susy", and Edward in 1556. She was often short-tempered with her daughter, and disliked Tilly's friend, Tom. Once Tilly was older, she found her mother "much easier to get on with."
Thomas "Tom" (born c. 1539) was the son of John the Carter and a friend of Tilly Middleton, though they sometimes went months without talking or seeing each other. He worked at the Royal Menagerie near the Tower of London. His dream was to become the Keeper of the Royal Menagerie. Tilly's parents and her friend, Frances Lea, disapproved of her seeing Tom. He was also friends with the Middleton's servant, Sal. By 1558, him and Tilly had grew apart and rarely saw each other.
- Anne Boleyn was the mother of Elizabeth. Shortly before her execution, she entrusted her diary, which contained a letter for Elizabeth, to Master Middleton.
- Edward VI (1537 – July 6, 1553) was the King of England. Before passing away, he declared his heir to be his cousin, Lady Jane Grey. He was the son of the previous King, Henry VIII.
- Doctor Feckenham was Queen Mary's confessor. He was sent to Lady Jane Grey by Mary to "save her soul." He tried converting her for several days, but Jane remained loyal to her religion.
- Gardiner, a former prisoner of the Tower. He was freed and appointed as Lord Chancellor by Mary.
- Guilford Dudley (died February 12, 1554) was the husband of Lady Jane Grey and son of the Duke of Northumberland. Him and his wife were executed, after the failed attempt to make Jane queen.
- Edward, (born c. 1556) Harry, (born c. 1553) and Jack Middleton (born c. 1550), Tilly's younger siblings, whom she often looked after. The youngest was named after his brother, who named at the age of two.
- Lady Jane Grey (1537 – February 12, 1554) was the cousin of Edward VI, Mary I, and Elizabeth. She inherited the throne from Edward, but ruled for only nine days. Mary subsequently became Queen and had Jane executed, though many thought she would be pardoned.
- Bishops Latimer and Ridley, two loyal Protestants tried for heresy and burned at the stake.
- Master Lea was Frances Lea's uncle. He worked at the Mint, also located in the Tower of London.
- Mary and Susannah "Susy" Middleton (born August 1554) were the younger twin sisters of Tilly.
- Duke of Northumberland (died August 22, 1553) was the father of Guilford Dudley. He unsuccessfully supported his daughter-in-law, Jane Grey to be Queen. The Duke was subsequently executed.
- Master Patridge was the Gentleman Gaoler of the Tower of London. Jane Grey was moved to his lodgings from the White Tower.
- Philip II was a Prince of Spain and the son of Charles V, the Holy Roman Emperor. Shortly becoming King of England, his father gave him the kingdoms of Naples and Jerusalem. In July 1544, he married Mary. He was unhappy with his marriage and often visited the Netherlands.
- Sal was the maidservant of the Middleton family. She competed with Tilly for Tom's attention, though Tilly eventually gave up.
- Duke of Suffolk (died February 23, 1554) was the father of Jane Grey. The Duke was imprisoned, after his daughter's short reign. Mary let him go free, but he was arrested again and executed.
- Susannah Middleton (died June 1553) was the younger sister of Tilly. She died shortly after being baptized. Her sibling, Susy, was named after her.
- Thomas Cranmer, the former Archbishop of Canterbury. He was imprisoned at the Tower of London. Cranmer was later burned at the stake for heresy.
- Thomas Wyatt was the leader of an uprising against Queen Mary's marriage to Philip of Spain. He surrendered and was imprisoned at the Tower, until his execution in April 1554.
- Unnamed was Tilly's uncle. He worked as the Chief Yeoman Warder.
- William Middleton was Tilly's older brother. He was his father's apprentice and acted very smug. As they grew older, William and Tilly were nicer to each other. He developed a crush on Tilly's friend, Frances.
- Master Worsley was the Keeper of the Royal Menagerie. He was often in a bad mood.