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Randolph Jefferson (October 1, 1755 – August 7, 1815) was the younger brother of Thomas Jefferson, the only male sibling to survive infancy. He was a planter and owner of the Snowden plantation that he inherited from his father. He served the local militia for about ten years, making captain of the local militia in 1794. He also served during the Revolutionary War. Randolph, known as "Uncle Randolph" when he visited Monticello, was considered as a candidate for the father of Sally Hemings's children following DNA studies that found that the Hemings children descended from the Jefferson line. The theory that Randolph Jefferson fathered Hemings children is discounted by the Thomas Jefferson Foundation given that records do not show that Randolph often visited Monticello. He often socialized with the enslaved people during his visits. His son, Isham Randolph Jefferson, who lived at Monticello during his childhood is another alternate candidate for Hemings children's paternity. Thomas Jefferson, though, was found by The Monticello Jefferson Hemings Report (2000) to be the likely father of Sally Hemings' children. Other scholars contend otherwise and find Randolph an attractive candidate.

Born at Shadwell, the Jefferson family plantation in Albemarle County, Virginia, his parents were Peter Jefferson, who died when Randolph was two years old, and Jane Randolph Jefferson. In 1764 and 1765, Randolph Jefferson studied with Ben Snead at the residence of his uncle Charles Lewis, Jr. and aunt Mary Randolph Lewis at Buck Island, which was a 960 acre tract located near Monticello and the Rivanna River in Albemarle County. He lived again at Shadwell with his mother in 1769, when he was taught by Patrick Morton. In 1770, the main house at Shadwell was destroyed in a fire, and his mother, Jane Randolph Jefferson, had a house built there as a replacement. He left Shadwell for Williamsburg when he was 16 He attended The Grammar School at the College of William & Mary and was tutored in higher subjects by Thomas Gwatkin, who taught mathematics and natural philosophy at the college. Additionally, he took violin lessons from Frances Alberti, as did his brother. Randolph Jefferson continued to “fiddle” throughout his life and willed his violin to his son, Robert Lewis Jefferson.

Thomas Jefferson described Randolph posthumously in a deposition that was taken as Randolph's sons contested the will that favored their stepmother, Mitchie Pryor Jefferson. Thomas was considerate and affectionate toward Randolph; they addressed each other as "Dear Brother," and exchanged visits and services with each other. Letters document that Thomas lent Randolph the harness for a gig, had his watch repaired, gave him a dog, sent him vegetable seeds, and gave him a spinning jenny. At Monticello, he was called "Uncle Randolph". A former Monticello enslaved man, Isaac Jefferson, recalled in 1847 that Randolph "used to come out among black people, play the fiddle and dance half the night "

Historian Dumas Malone states that Randolph did not share his older brother's eloquence. His letters to Thomas show a disregard of grammar and the use of colloquialisms such as "tech" instead of "touch." He was a member of the local militia in 1779. Along with his brother, Jefferson signed an Oath of Allegiance to the Commonwealth of Virginia in 1777, He, his brother, and Charles Lewis also signed the Albemarle County Oath of Allegiance to the Commonwealth on April 21, 1779. It was also called the Albemarle Declaration of Independence. During the Revolutionary War (1775–1783), he served under General Thomas Nelson with the Virginia Light Dragoons. In the fight against Tarleton, in the summer of 1781, he provided provisions for the Virginia troops, volunteered a slave from Snowden to help move items from military stores at Scotts Ferry in Albemarle County to Bedford County, and allowed the 3rd Regiment of Light Dragoons to camp at Snowden for over a month and a half. Following the war, Jefferson served in the Buckingham County Militia, achieving the rank of Lieutenant in 1787 and, in 1794, was recommended to rank of Captain. After at least nine years of service in the Militia, Randolph's reason for not continuing remains a mystery. He was content to enjoy the title of Captain the remainder of his life.

In 1776, Randolph inherited the Snowden plantation in Buckingham County, Virginia. with 2,291⅔ acres called "Fluvanna lands" located near the Hardware River and Scottsville,|name=acres from his father, Peter Jefferson's estate. More specifically, the plantation was located along the James River, about twenty miles south of Monticello and across from Scott's Ferry and on the south side of Horseshoe Bend. Months after Randolph's death, the dwelling house at Snowden burned to the ground. Ultimately, none of Randolph Jefferson's sons could afford to purchase Snowden from their father's estate and it was sold to Capt. John Harris of Albemarle County. Most of them, eventually settled nearby in Scottsville, Albemarle and Fluvanna County, Virginia.

On July 30, 1781, Jefferson married his first cousin, Anne Lewis, the daughter of Colonel Charles Lewis of Buck Island and Mary Randolph, the sister of Jane Randolph Jefferson. Robert Lewis, Peter Field, Isham Randolph, James Lilburne and Anna Scott. Randolph was a widower for about ten years after his wife died about 1799. Mitchie, whose father was David Pryor, — but there was a match to the Jefferson male line to descendants of Sally Hemings' son Eston. led to speculation by deniers of the scientific evidence about whether Randolph was the Jefferson who fathered the Hemings children. The Thomas Jefferson Heritage Society, a fringe group formed in 1999 to "defend the memory of Thomas Jefferson", commissioned its own independent scholars' report that was completed in 2001. While the report, without evidence, suggested that Randolph Jefferson, or one of his sons, could have fathered Hemings's children, it is more likely that Thomas Jefferson was the father. Hemings said that "Jefferson" was the father of his children. William G. Hyland, Jr. asserts, without foundation, that Jefferson could refer to Randolph Jefferson, who lived 20 miles from Monticello and socialized with the slaves there only on the basis that he had the same Y DNA as his brother, Thomas. Eston Hemings family believed that their ancestor was a "Jefferson uncle". There are no entries in Thomas Jefferson's record books for Isham. According to the study of the paternity of Sally Hemings's children: "As mentioned elsewhere, no one familiar with Monticello suggested that Sally Hemings was promiscuous or that her children had multiple fathers."