Next-generation sequencing of multiple individuals per barcoded library by deconvolution of sequenced amplicons using endonuclease fragment analysis

TY - JOUR

T1 - Next-generation sequencing of multiple individuals per barcoded library by deconvolution of sequenced amplicons using endonuclease fragment analysis

AU - Andersen, Jeppe D

AU - Pereira, Vania

AU - Pietroni, Carlotta

AU - Mikkelsen, Martin

AU - Johansen, Peter

AU - Børsting, Claus

AU - Morling, Niels

PY - 2014/8

Y1 - 2014/8

N2 - The simultaneous sequencing of samples from multiple individuals increases the efficiency of next-generation sequencing (NGS) while also reducing costs. Here we describe a novel and simple approach for sequencing DNA from multiple individuals per barcode. Our strategy relies on the endonuclease digestion of PCR amplicons prior to library preparation, creating a specific fragment pattern for each individual that can be resolved after sequencing. By using both barcodes and restriction fragment patterns, we demonstrate the ability to sequence the human melanocortin 1 receptor (MC1R) genes from 72 individuals using only 24 barcoded libraries.

AB - The simultaneous sequencing of samples from multiple individuals increases the efficiency of next-generation sequencing (NGS) while also reducing costs. Here we describe a novel and simple approach for sequencing DNA from multiple individuals per barcode. Our strategy relies on the endonuclease digestion of PCR amplicons prior to library preparation, creating a specific fragment pattern for each individual that can be resolved after sequencing. By using both barcodes and restriction fragment patterns, we demonstrate the ability to sequence the human melanocortin 1 receptor (MC1R) genes from 72 individuals using only 24 barcoded libraries.

U2 - 10.2144/000114200

DO - 10.2144/000114200

M3 - Journal article

C2 - 25109295

SN - 0736-6205

VL - 57

SP - 91

EP - 94

JO - BioTechniques

JF - BioTechniques

IS - 2

ER -