Template Prep

Please note that the recommended quantities are TOTAL template amounts. The Sanger facility uses 4ul of template/sequencing reaction. For requests with standard size plasmids, we recommend new users start by submitting 10ul of ~50ng/ul plasmid and 2ul of 10uM primer (for a total volume of 12ul).  If you are requesting one of the core primers we offer, then you only need to submit your template. Please note that each primer-template combination may need to be optimized to generate good quality data. For customer cleaned PCR products, we recommend submitting 10ul of 0.5-2ng/ul template + 2ul 10uM primer. If you are submitting PCR reactions for the facility to clean up, we recommend beginning with ~50ng of PCR product in a volume of 20ul along with a separate aliquot of at least 3ul/sequencing reaction of 10uM primer.  If you have questions or need help with troubleshooting, please email sanger@utgsaf.org.