改进的化学发光DNA测序法。
Improved chemiluminescent DNA sequencing.
作者信息
Martin C, Bresnick L, Juo R R, Voyta J C, Bronstein I
机构信息
Tropix, Inc., Bedford, MA 01730.
出版信息
Biotechniques. 1991 Jul;11(1):110-3.
A new chemiluminescent 1,2-dioxetane substrate, CSPD, shows improved performance over AMPPD when used in our nonisotopic method for DNA sequencing. CSPD differs from AMPPD by the addition of a chlorine atom to the adamantyl group that limits the amount of aggregation of the dioxetane and its dephosphorylated anion. This results in a shorter time elapsing before reaching steady state light emission when detecting nucleic acids on nylon membrane. An additional advantage of CSPD over AMPPD is that the resolution of imaged DNA bands does not degrade over time. These features of CSPD permit rapid acquisition of high-quality DNA sequence data.
一种新型化学发光1,2 - 二氧杂环丁烷底物CSPD，在我们用于DNA测序的非同位素方法中使用时，比AMPPD表现出更好的性能。CSPD与AMPPD的不同之处在于，在金刚烷基上添加了一个氯原子，这限制了二氧杂环丁烷及其去磷酸化阴离子的聚集量。这导致在尼龙膜上检测核酸时，达到稳态发光前经过的时间更短。CSPD相对于AMPPD的另一个优点是，成像的DNA条带分辨率不会随时间下降。CSPD的这些特性使得能够快速获取高质量的DNA序列数据。
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