- Dorothea Brooke: An intelligent, wealthy woman with great aspirations, Dorothea avoids displaying her wealth and embarks upon projects such as redesigning cottages for her uncle's tenants. She marries the elderly Reverend Edward Casaubon, with the idealistic idea of helping him in his research, The Key to All Mythologies. However, the marriage was a mistake, as Casaubon fails to take her seriously and resents her youth, enthusiasm, and energy. Her requests to assist him make it harder for him to conceal that his research is years out of date. Faced with Casaubon's coldness on their honeymoon, Dorothea becomes friends with his relative, Will Ladislaw. Some years after Casaubon's death she falls in love with Will and marries him.
- Tertius Lydgate: An idealistic, talented, but naive young doctor, is relatively poor, but of good birth. He hopes to make big advances in medicine through his research, but ends up in an unhappy marriage with Rosamond Vincy. His attempts to show he is answerable to no man fail, and he eventually has to leave town, sacrificing his high ideals to please his wife.
- Rev. Edward Casaubon /ˈkæzəbɑːn/: A pedantic, selfish, elderly clergyman who is so taken up with his scholarly research that his marriage to Dorothea is loveless. His unfinished book, The Key to All Mythologies, is intended as a monument to Christian syncretism, but his research is out of date as he cannot read German. He is aware of this but admits it to no one.
- Mary Garth: The plain, kind daughter of Caleb and Susan Garth serves as Mr Featherstone's nurse. She and Fred Vincy were childhood sweethearts, but she will not let him woo her until he shows himself willing and able to live seriously, practically and sincerely.
- Arthur Brooke: The oft-befuddled, none-too-clever uncle of Dorothea and Celia Brooke has a reputation as the worst landlord in the county, but stands for Parliament on a Reform platform.
- Celia Brooke: Dorothea's younger sister is a beauty. She is more sensual than Dorothea and does not share her idealism and asceticism. She is only too happy to marry Sir James Chettam when Dorothea rejects him.
- Sir James Chettam: A neighbouring landowner, he is in love with Dorothea and helps with her plans to improve conditions for the tenants. When she marries Casaubon, he marries Celia Brooke.
- Rosamond Vincy: Vain, beautiful and shallow, Rosamond has a high opinion of her own charms and a low opinion of Middlemarch society. She marries Tertius Lydgate, believing he will raise her social standing and keep her comfortable. When her husband meets financial difficulties, she thwarts his efforts to economise, seeing such sacrifices as beneath her and insulting. She cannot bear the idea of losing social status.
- Fred Vincy: Rosamond's brother has loved Mary Garth from childhood. His family hopes he will advance socially by becoming a clergyman, but he knows Mary will not marry him if he does. Brought up to expect an inheritance from his uncle, Mr Featherstone, he is a spendthrift, but later changes through his love for Mary and finds by studying under Mary's father a profession that gains Mary's respect.
- Will Ladislaw: This young cousin of Mr Casaubon has no property, as his grandmother married a poor Polish musician and was disinherited. He is a man of verve, idealism and talent, but no fixed profession. He is in love with Dorothea, but cannot marry her without her losing Mr Casaubon's property.
- Humphrey Cadwallader and Elinor Cadwallader: Neighbours of the Brookes, Mr Cadwallader is a rector and Mrs Cadwallader a pragmatic and talkative woman who comments on local affairs with wry cynicism. She disapproves of Dorothea's marriage and Mr Brooke's parliamentary endeavours.
- Walter Vincy and Lucy Vincy: A respectable manufacturing couple, they wish their children to advance socially and are disappointed by Rosamond's and Fred's marriages. Vincy's sister is married to Nicholas Bulstrode. Mrs Vincy was an innkeeper's daughter and her sister the second wife of Mr. Featherstone.
- Caleb Garth: Mary Garth's father is a kind, honest, generous surveyor and land agent involved in farm management. He is fond of Fred and eventually takes him under his wing.
- Camden Farebrother: A poor but clever vicar and amateur naturalist, he is a friend of Lydgate and Fred Vincy and loves Mary Garth. His position improves when Dorothea appoints him to a living after Casaubon's death.
- Nicholas Bulstrode: A wealthy banker married to Vincy's sister, Harriet, he is a pious Methodist keen to impose his beliefs in Middlemarch society. However, he has a sordid past he is desperate to hide. His religion favours his personal desires and lacks sympathy for others.
- Peter Featherstone: An old landlord of Stone Court, he is a self-made man, who has married Caleb Garth's sister. On her death he takes Mrs Vincy's sister as his second wife.
- Jane Waule: A widow and Peter Featherstone's sister, she has a son, John.
- Mr Hawley: A foul-mouthed businessman, he is an enemy of Bulstrode.
- Mr Mawmsey: A grocer
- Dr Sprague: A Middlemarch physician
- Mr Tyke: A clergyman favoured by Bulstrode
- Joshua Rigg Featherstone: Featherstone's illegitimate son, he appears at the reading of Featherstone's will and receives a fortune instead of Fred. He is also the stepson of John Raffles, who comes into town to visit Rigg, but instead reveals Bulstrode's past. His appearance in the novel is crucial to the plot.
- John Raffles: Raffles is a braggart and a bully, a humorous scoundrel in the tradition of Sir John Falstaff, and an alcoholic. But unlike Falstaff, Raffles is a truly evil man. He holds the key to Bulstrode's dark past and Lydgate's future.
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