Size-Selection of Cell-Free DNA for Increasing Family Size During Next-Generation Sequencing

Abstract

A method of increasing detection of low-abundant fragments of cell-free DNA (ccfDNA) in a biological sample is disclosed and discussed. Such a method can include isolating an initial fraction of ccfDNA fragments from a biological sample, ligating a unique molecular identifier (UMI) to each of the ccfDNA fragments in the initial fraction, amplifying the plurality of ccfDNA fragments to generate a ccfDNA library, isolating a short fraction of ccfDNA fragments from the ccfDNA library, where the ccfDNA fragments in the short fraction are limited to a size of less than or equal to 160 base pairs (bp), amplifying the ccfDNA fragments in the short fraction, and sequencing the ccfDNA fragments in the short fraction to generate sequenced ccfDNA fragments.

1 . A method of increasing detection of low-abundant fragments of cell-free DNA (ccfDNA) in a biological sample from a subject, comprising:
 isolating an initial fraction of ccfDNA fragments from a biological sample;   ligating a unique molecular identifier (UMI) to each of the ccfDNA fragments in the initial fraction;   amplifying the plurality of ccfDNA fragments to generate a ccfDNA library;   isolating a short fraction of ccfDNA fragments from the ccfDNA library, where the ccfDNA fragments in the short fraction are limited to a size of less than or equal to 160 base pairs (bp);   amplifying the ccfDNA fragments in the short fraction; and   
       sequencing the ccfDNA fragments in the short fraction to generate sequenced ccfDNA fragments.