w6.1.02 | c6.
PROD | u6.1.6


The truth is not always what it appears to be

Restriction sites other than in MCS


This is a fact, indeed. You only need to cut your vector of choice with one or two suitable enzymes of the MCS. Your gene insert can then be PCR amplified with primers that generate homology to the vector at the ends of the PCR product. These homologous regions can then be used for subcloning via SLIC (= sequence and ligation independent cloning) into your vector. Any gene internal restriction sites do not matter at all because the PCR product won’t be cut during subcloning. Eurofins’ molecular biology experts will do the design and make sure that all your requirements are met, e.g., in frame cloning. SLIC is as fast and efficient as traditional subcloning via restriction enzyme sites and, if you have forgotten, e.g., to introduce a stop codon or a fusion tag at the end of your gene, this can also be fixed simultaneously.

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