Pekarek was born and raised in Denver, Colorado.
She grew up in a household with her parents and one sibling, an older sister.
She began playing the cello at age nine, performing in her school's orchestra.
She chose the cello because the school's policy was that only fifth-graders could play woodwind instruments, while fourth-graders could play stringed instruments.
Explaining her thinking, she said, "...I went with cello because I wanted to get started sooner."
Growing up, she enjoyed listening to powerful vocalists such as Etta James, Ella Fitzgerald, Aretha Franklin, and Otis Redding.
Her parents introduced her to American folk music via vinyl records of Bob Dylan, Carole King, and Emmylou Harris.
She attended college at the University of Northern Colorado, where she initially majored in vocal performance and musical theatre.
She changed majors, however, graduating with a degree in music education and planning to become a choir teacher.
Although she did not play the cello much while attending college, she earned some income post-graduation as a cellist at weddings.
After finishing her degree in music education, Pekarek responded to a Craigslist ad posted by Wesley Schultz and Jeremiah Fraites. Schultz and Fraites were looking for a cellist to join their band, The Lumineers. Explaining why she responded to the ad, Pekarek said, "I was living with my parents and kind of just looking for something to do. I thought it would be a fun way to pass my time until I found something full time to do."
For several months, the three rehearsed and played shows around Denver. They decided to undertake a small-scale concert tour for 30 days, playing shows from Portland, Oregon to New York City. Of her decision to tour with the band, Pekarek said, "I was only 23 years old. If you are going to be in a band and do something stupid, it might as well be then." The 30-day tour was followed by a tour of the West Coast and a weekly installment at The Living Room, a venue in New York.

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