Robbins C M, Hsu E, Gillevet P M
National Center for Human Genome Research, National Institutes of Health, Bethesda, MD 20892-4470, USA.
Biotechniques. 1996 May;20(5):862-4, 866-8. doi: 10.2144/96205st06.
We investigated the use of Taq dye primer and Taq terminator sequencing chemistry to optimize the quality of sequence data obtained from templates containing homopolymer tracts and repetitive elements. In direct side-by-side comparisons using the Applied Biosystems Model 373A Fluorescent Sequencer, the Taq terminator sequencing chemistry gave much cleaner and more consistent results on long homopolymer tracts and dinucleotide repeats. We also investigated various thermal cycling conditions and determined that higher annealing temperatures and longer denaturation times improved the ability to sequence through these problem templates.
我们研究了使用Taq染料引物和Taq终止子测序化学方法来优化从包含同聚物片段和重复元件的模板中获得的序列数据的质量。在使用应用生物系统公司373A荧光测序仪进行的直接并排比较中,Taq终止子测序化学方法在长同聚物片段和二核苷酸重复序列上产生了更清晰、更一致的结果。我们还研究了各种热循环条件,并确定较高的退火温度和较长的变性时间提高了通过这些有问题的模板进行测序的能力。