William Bradford's Of Plymouth Plantation was written over the course of two decades between 1630 and 1650. It was published in 1898 and is widely regarded as a the single best piece of evidence for the lifestyle of the pilgrims and their settlement. The characters are many, as the narration comprises twenty years of living in the Plymouth colony.

The major characters include Isaac Allerton, who was first appointed as an assistant to William Bradford. He eventually goes to England to advocate on behalf of the colonists. He was closely involved in the finances of the colonists and paid debts on their behalf. Eventually, however, he mixes the colonists finances with his own. He incurs debts on their behalf and acts as an independent merchant.

Another major (though relatively short-lived) character, is Robert Cushman. His family was from Leiden, and Cushman acts as a chief agent on behalf of the separatists. He comes to the New World in 1621 to convince the colonists to sign a contract from London's Virginia company. According to Bradford, Cushman came back to Plymouth to spend his last days before dying.

Myles Standish is another primary character in Bradford's account. He set sail from England on the Speedwell, whose crew in turn joined the Mayflower at Southampton. He is one of few to survive the journey. Myles Standish brings his military expertise to the settlement, and serves as a strong leader for the disillusioned settlers.

Squanto is a unique figure and important liaison between the colonists and Native Americans. He had been kidnapped and spent time in England, where he perfected the English language. He is friendly with the English settlers, and when he was taken prisoner by a Sachem at Namasket, the pilgrims rescued him.

A final important character is John Winthrop, a close friend to William Bradford and governor of Massachusetts. He was a Puritan and was rather conservative in his administration. He was instrumental in warding off the threat of the Pequots and ultimately waged the Pequot War.