Primer walking

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Primer walking determines the sequences of PCR products or plasmids that are not analyzable by one single primer extension. Internal primers are designed and prepared with results obtained after performing end sequencing using client-provided or designed primers.

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This service works by continuous extension through designing an internal primer at an appropriate point using the result obtained from the reaction conducted with a new primer and the same template. For lengths of more than 15 kb, large construct DNA shotgun sequencing is recommended.

This service reads 2~15 kb of sequence of PCR products or plasmid DNA that cannot be read fully by single primer extension sequencing. Once performing end sequencing with designated primer by customer, we design internal primers with reference to end sequence data. Necessary walking continues with the same method until the whole region is covered. It takes around one week to read 1~1.5 kb by primer walking.

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