Unknown

Dataset Information

0

Multichange isothermal mutagenesis: a new strategy for multiple site-directed mutations in plasmid DNA.


ABSTRACT: Multichange ISOthermal (MISO) mutagenesis is a new technique allowing simultaneous introduction of multiple site-directed mutations into plasmid DNA by leveraging two existing ideas: QuikChange-style primers and one-step isothermal (ISO) assembly. Inversely partnering pairs of QuikChange primers results in robust, exponential amplification of linear fragments of DNA encoding mutagenic yet homologous ends. These products are amenable to ISO assembly, which efficiently assembles them into a circular, mutagenized plasmid. Because the technique relies on ISO assembly, MISO mutagenesis is additionally amenable to other relevant DNA modifications such as insertions and deletions. Here we provide a detailed description of the MISO mutagenesis concept and highlight its versatility by applying it to three experiments currently intractable with standard site-directed mutagenesis approaches. MISO mutagenesis has the potential to become widely used for site-directed mutagenesis.

SUBMITTER: Mitchell LA 

PROVIDER: S-EPMC4040258 | BioStudies | 2013-01-01

REPOSITORIES: biostudies

Similar Datasets

1000-01-01 | S-EPMC4333370 | BioStudies
1000-01-01 | S-EPMC5789069 | BioStudies
2008-01-01 | S-EPMC2629768 | BioStudies
2019-10-03 | GSE87804 | GEO
2019-10-03 | GSE87804 | GEO
2008-01-01 | S-EPMC7419347 | BioStudies
2019-01-01 | S-EPMC6343910 | BioStudies
2015-01-01 | S-EPMC4459934 | BioStudies
2000-01-01 | S-EPMC108465 | BioStudies
2017-01-01 | S-EPMC5615473 | BioStudies