Cycle Sequencing

There are several brands of fluorescent dye kits available that incorporate fluorescent dideoxynucleotides into extension products.Each kit basically comes with Taq DNA Polymerase, fluorescent-ddNTPs, & dNTPs.
There are four different fluorescent dyes that distinguish between the four bases in DNA. The reaction is linear; only one primer is used per reaction. The LGT set up the sequencing procedure based on the ABI BigDye Terminator chemistry. Refer to LGT Cycle Sequencing Protocol site or the ABI protocol for more information on preparing reactions. The LGT has three ABI 2700 and four Eppendorf MasterCycler PCR machines available for cycle sequencing reactions. The cycle sequencing run time is about 2.5 hours. See lab manager for more information.

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