All samples sent to the Genomics Core for Sanger sequencing should be supplied in water, rather than buffer. EDTA inhibits polymerase activity. We prefer all samples to be measured here in the lab using one of our fluorometers for consistency. There is no cost for you to measure your own samples, or we can measure your samples for you for a small fee.
All primers should be delivered in water at 10 uM concentration, 2 ul per reaction.
For DNA templates we use 15ngs DNA per 100 base pairs in length in 12 ul volumes or less.
For example, a 5 kb plasmid requires 750 ng of template DNA. For a 500 bp PCR product, 75 ng of template are required.