Effects of Agarose and LB Medium on Dye-terminator DNA Sequencing

Inhibitory effects of agarose gel and LB medium on DNA sequencing.

The direct sequencing of PCR products, a bacterial colony (plasmid DNA), or phage plaque (λDNA) is a very powerful technique applicable to several molecular biological experiments. In particular, the sequencing of plasmid DNA directly from within bacterial colonies provides a rapid screening method to confirm the orientation of DNA inserts. Similarly, the direct sequencing of λDNA within plaque...

Direct colony sequencing of plasmid DNA by dye terminator methods.

Direct colony sequencing of plasmid DNA is a powerful technique. Radioactive or fluorescent dye primer methods are the reported direct colony sequencing methods (2–4,6). Recently, dye terminator sequencing chemistry (7) has been improved by the introduction of a modified enzyme, AmpliTaq DNA polymerase FS (8), as well as a new d-Rhodamine and BigDye terminator (PE Biosystems, Foster City, CA,...

Anchored oligo(dT) primers for automated dye terminator DNA sequencing.

AmpliTaq DNA polymerase, FS dye-terminator sequencing: analysis of peak height patterns.

Taq DNA polymerases in which the phenylalanine is substituted by a tyrosine at position 667 (Taq F667Y) are members of a new class of DNA polymerases that incorporate chain-terminating dideoxyribonucleoside triphosphates (ddNTPs) much more efficiently than the wild-type Taq DNA polymerase. Improved incorporation of ddNTPs into DNA during cycle sequencing using AmpliTaq DNA polymerase, FS (Taq-F...

Octamer-primed cycle sequencing using dye-terminator chemistry.

Octamer Sequencing Technology, OST, is a method of DNA sequencing using single octamer oligonucleotides to prime cycle sequencing reactions. This sequencing strategy is faster than a traditional primer-walking strategy, since access to this optimized octamer library eliminates delays associated with designing and synthesizing gene specific primers. In this report, OST has been optimized for flu...