Improved next-generation sequencing pre-capture library yields and sequencing parameters using on-bead PCR.
Biotechniques
; 68(1): 48-51, 2020 01.
Article
em En
| MEDLINE
| ID: mdl-31825238
ABSTRACT
Tumor DNA sequencing results can have important clinical implications. However, its use is often limited by low DNA input, owing to small tumor biopsy size. To help overcome this limitation we have developed a simple improvement to a commonly used next-generation sequencing (NGS) capture-based library preparation method using formalin-fixed paraffin-embedded-derived tumor DNA. By using on-bead PCR for pre-capture library generation we show that library yields are dramatically increased, resulting in decreased sample failure rates. Improved yields allowed for a reduction in PCR cycles, which translated into improved sequencing parameters without affecting variant calling. This methodology should be applicable to any NGS system in which input DNA is a limiting factor.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Biblioteca Gênica
/
Reação em Cadeia da Polimerase
/
Sequenciamento de Nucleotídeos em Larga Escala
Limite:
Humans
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Austrália