A prolific and robust whole-genome genotyping method using PCR amplification via primer-template mismatched annealing.
J Integr Plant Biol
; 65(3): 633-645, 2023 Mar.
Article
em En
| MEDLINE
| ID: mdl-36269601
ABSTRACT
Whole-genome genotyping methods are important for breeding. However, it has been challenging to develop a robust method for simultaneous foreground and background genotyping that can easily be adapted to different genes and species. In our study, we accidently discovered that in adapter ligation-mediated PCR, the amplification by primer-template mismatched annealing (PTMA) along the genome could generate thousands of stable PCR products. Based on this observation, we consequently developed a novel method for simultaneous foreground and background integrated genotyping by sequencing (FBI-seq) using one specific primer, in which foreground genotyping is performed by primer-template perfect annealing (PTPA), while background genotyping employs PTMA. Unlike DNA arrays, multiple PCR, or genome target enrichments, FBI-seq requires little preliminary work for primer design and synthesis, and it is easily adaptable to different foreground genes and species. FBI-seq therefore provides a prolific, robust, and accurate method for simultaneous foreground and background genotyping to facilitate breeding in the post-genomics era.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Genoma
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article