Construction of a 15-plex Rapid STR Multiplex Amplification System / 法医学杂志
Journal of Forensic Medicine
; (6): 49-53, 2016.
Article
em Zh
| WPRIM
| ID: wpr-984042
Biblioteca responsável:
WPRO
ABSTRACT
OBJECTIVE@#To establish a 15-plex rapid STR multiplex amplification system.@*METHODS@#Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives.@*RESULTS@#Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure.@*CONCLUSION@#The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
DNA
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Reação em Cadeia da Polimerase
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Impressões Digitais de DNA
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Mapeamento Cromossômico
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Sensibilidade e Especificidade
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Repetições de Microssatélites
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Sequências de Repetição em Tandem
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Grupos Raciais
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Alelos
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Genética Forense
Limite:
Humans
Idioma:
Zh
Revista:
Journal of Forensic Medicine
Ano de publicação:
2016
Tipo de documento:
Article