Ready-to-run sequencing is an option for those who prefer to do sequencing reactions and purifications themselves.
- Sequencing reactions done with BigDyeTerminator v1.1 cycle sequencing kit (Applied Biosystems)
- Ready-to-run samples are brought in dried.
Template purification i.e. removal of PCR reaction “residuals” (primers, dNTPs, salt, non-specific PCR products) is crucial for good quality sequence. Depending on the quality of the PCR product methods that can be used are:
- ExoI/SAP treatment
- Commercial column purification
- Gel purification
- Purification with magnetic beads (Agencourt Ampure or similar)
Last updated: 7/1/2016