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Old 05-10-2013, 12:23 PM   #2
Senior Member
Location: Boston,MA

Join Date: Nov 2008
Posts: 122

You need to design a custom sequencing primer.

Use the sequence of the adapter+sequence of your non-variable region(10 bases)

Then create a sequencing primer that is complementary to the last base of your nonvariable region-then work back.

When the Tm of the primer is above 65 (that way it will work on both HiSeq and MiSeq) you have that primer synthesized and HPLC purified.

Use the primer at 100uM.

Whoever is doing you sequencing will know how to use it.
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