Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers

ID: 79

Group:

Methods for DNA sequencing

Prepared by:

Bruce A. Roe (broe@uoknor.edu)

Detail:

Each base-specific fluorescent-labeled cycle sequencing reaction routinely included approximately 100 or 200 ng Biomek isolated single-stranded DNA for A and C or G and T reactions, respectively. Double-stranded cycle sequencing reactions similarly contained approximately 200 or 400 ng of plasmid DNA, isolated using either the standard alkaline lysis or the diatomaceous earth modified alkaline lysis procedures. All reagents except template DNA are added in one pipetting step from a premix of previously aliquotted stock solutions stored at -20degC (see Appendix B).

Year prepared: NULL

Date uploaded: 2010-02-26 14:28:18

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