You say your machines can read sequences up to about 800 nt. What if my DNA is longer?

How can I determine the sequence for the whole thing?

The U of M DNA Sequencing Core provides only raw sequencing. If you provide a template and a primer, we will return the results of *one* sequencing reaction (typically 800+nt for good templates). **** We do not perform primer-walking or shotgun sequencing. **** If you need the sequence of a larger fragment, please see the following flowchart to determine your options.

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If you need a complete sequence from a larger fragment, here's how to proceed:

• Are you looking for mutations in a known sequence?

  We recommend that you design primers all at once that will cover the entire length of your insert. See our web page on designing sequencing primers. For recommendations on the placement of those primers, please see the section on Primer Walking. Have all your primers synthesized at once and submit them to the Core for simultaneous sequencing.

  We would be happy to assemble the final sequence if you want; please see Assembly of Sequencing Data for discussion of software and techniques for that step.

• Is your insert less than 5 kb in length?

  We recommend Primer Walking to get your complete sequence. The Core can perform only some of the steps necessary in this procedure (click the link Primer Walking to learn more). You may also want to see our discussion on designing sequencing primers.

• You have a cloned fragment of more than 5 kb.

  We recommend shotgun sequencing to obtain your complete sequence. The Core can perform only some of the steps of this procedure (click the link to learn more).

Contact Dr. Robert Lyons, the Core Director, with questions or suggestions.
21-Jun-01