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Center for Protein and Nucleic Acid Research

Sequencing of GC Rich/Secondary Structure Template
(>65% GC)

We do not guarantee results for templates of this type, although we can use special methods to try to alleviate the problem (note: primers located too close to the secondary structure may not prime and give a blank/failed reactions).

Our first method of choice is a straight chemical manipulation of the sequencing reaction. We add DMSO to the sample, which seems to aid in denaturing the sample.

If this fails to work, we can try a second procedure utilizing DNA single-strand binding protein.

If neither of these methods work we recommend that you try linearizing your sample or submitting the PCR product. This may or may not alleviate the problem.

Making your template single stranded is usually the most foolproof method for getting good sequences results on these templates. We try to avoid asking you to do this because we are aware of the work it would require of you. But sometimes it can be necessary according to your data needs.

Each time we run a sequencing reaction, usual sequencing charges will be applied.