Bowers Jayson, Mitchell Judith, Beer Eric, Buzby Philip R, Causey Marie, Efcavitch J William, Jarosz Mirna, Krzymanska-Olejnik Edyta, Kung Li, Lipson Doron, Lowman Geoffrey M, Marappan Subramanian, McInerney Peter, Platt Adam, Roy Atanu, Siddiqi Suhaib M, Steinmann Kathleen, Thompson John F
Helicos BioSciences Corporation, Cambridge, Massachusetts, USA.
Nat Methods. 2009 Aug;6(8):593-5. doi: 10.1038/nmeth.1354. Epub 2009 Jul 20.
We synthesized reversible terminators with tethered inhibitors for next-generation sequencing. These were efficiently incorporated with high fidelity while preventing incorporation of additional nucleotides, and we used them to sequence canine bacterial artificial chromosomes in a single-molecule system that provided even coverage for over 99% of the region sequenced. This single-molecule approach generated high-quality sequence data without the need for target amplification and thus avoided concomitant biases.
我们合成了带有连接抑制剂的可逆终止子用于下一代测序。这些可逆终止子能够高效且高保真地掺入，同时阻止额外核苷酸的掺入，并且我们使用它们在单分子系统中对犬类细菌人工染色体进行测序，该系统为超过99%的测序区域提供了均匀覆盖。这种单分子方法无需目标扩增即可生成高质量的序列数据，从而避免了相关的偏差。
