A new method has been developed for rapidly closing a large number of gaps in a whole-genome shotgun sequencing project. The method employs multiplex PCR and a novel pooling strategy to minimize the number of laboratory procedures required to sequence the unknown DNA that falls in between contiguous sequences. Multiplex sequencing, a novel procedure in which multiple PCR primers are used in a single sequencing reaction, is used to interpret the multiplex PCR results. Two protocols are presented, one that minimizes pipetting and another that minimizes the number of reactions. The pipette optimized multiplex PCR method has been employed in the final phases of closing the Streptococcus pneumoniae genome sequence, with excellent results.
Optimized Multiplex PCR: Efficiently Closing a Whole-Genome Shotgun Sequencing Project
최적화된 다중 PCR: 전체 유전체 샷건 시퀀싱 프로젝트 효율적으로 마무리하기
[Category] 폐렴구균 감염증,
[Article Type] journal-article
[Source] pubmed
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