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MICROFLUIDIC SEQUENCING TECHNIQUES

机译:微流控测序技术

摘要

The present invention generally relates to microfluidics and, in some embodiments, to the determination of cells. In some aspects, primers able to introduce restriction sites into certain amplified nucleic acids are used. For example, the primers may introduce restriction sites into normal (wild-type) nucleic acids, but be unable to introduce restriction sites into mutant nucleic acids, e.g., due to a mismatch in the nucleic acid sequences caused by the mutant. After amplification, the nucleic acids may be exposed to a suitable restriction enzyme, which may cleave normal nucleic acids but not the mutant nucleic acids. In this way, mutant nucleic acids may be relatively quickly identified. In some embodiments, cells may be contained within microfluidic droplets and assayed to determine the mutant cells. In certain cases, for example, the nucleic acids may be amplified within droplets and attached to suitable tags, e.g., prior to breaking or merging the droplets and sequencing of the nucleic acids.
机译:本发明通常涉及微流体,并且在一些实施方案中,涉及细胞的确定。在一些方面,使用能够将限制性位点引入某些扩增的核酸的引物。例如,引物可以将限制位点引入正常(野生型)核酸中,但是由于例如由突变体引起的核酸序列的错配,而不能将限制位点引入突变体核酸中。扩增后,可将核酸暴露于合适的限制酶,其可切割正常核酸,但不切割突变体核酸。以这种方式,可以相对快速地鉴定突变核酸。在一些实施方案中,细胞可被包含在微流液滴中并被测定以确定突变细胞。在某些情况下,例如,核酸可以在小滴内扩增并附着在合适的标签上,例如,在破坏或合并小滴并进行核酸测序之前。

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