Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
A General Strategy of Aerolysin Nanopore Detection for Oligonucleotides with the Secondary Structure
Small ( IF 13.0 ) Pub Date : 2018-03-30 , DOI: 10.1002/smll.201704520 Dong-Fang Liao 1 , Chan Cao 1 , Yi-Lun Ying 1 , Yi-Tao Long 1
Small ( IF 13.0 ) Pub Date : 2018-03-30 , DOI: 10.1002/smll.201704520 Dong-Fang Liao 1 , Chan Cao 1 , Yi-Lun Ying 1 , Yi-Tao Long 1
Affiliation
|
An aerolysin nanopore is employed as a sensitive tool for single‐molecule analysis of short oligonucleotides (≤10 nucleotides), poly(ethylene glycol) (PEGs), peptides, and proteins. However, the direct analysis of long oligonucleotides with the secondary structure (e.g., G‐quadruplex topology) remains a challenge, which impedes the further practical applications of the aerolysin nanopore. Here, a simple and applicable method of aerolysin nanopore is presented to achieve a direct analysis of structured oligonucleotides that are extended to 30 nucleotides long by a cation‐regulation mechanism. By regulating the cation type in electrolyte solution, the structured oligonucleotides are unfolded into linear form which ensures the successive translocation. The results show that each model oligonucleotide of 5′‐(TTAGGG)n‐3′ can produce a well‐resolved current blockade in its unfolded solution of MgCl2. The length between 6 and 30 nucleotides long of model oligonucleotides is proportional to the duration time, showing a translocation velocity as low as 0.70–0.13 ms nt−1 at +140 mV. This method exhibits an excellent sensitivity and a sufficient temporal resolution, provides insight into the aerolysin nanopore methodology for genetic and epigenetic biosensing, making aerolysin applicable in practical diagnosing with long and structured nucleic acids.
中文翻译：
溶血素纳米孔检测二级结构寡核苷酸的一般策略。
气溶素纳米孔被用作对短寡核苷酸（≤10个核苷酸），聚乙二醇（PEG），肽和蛋白质进行单分子分析的灵敏工具。但是，直接分析具有二级结构（例如，G-四链体拓扑）的长寡核苷酸仍然是一个挑战，这阻碍了溶血素纳米孔的进一步实际应用。在这里，提出了一种简单而适用的气溶素纳米孔方法，以实现对结构化寡核苷酸的直接分析，该结构化寡核苷酸通过阳离子调节机制延伸至30个核苷酸长。通过调节电解质溶液中的阳离子类型，结构化的寡核苷酸可展开为线性形式，从而确保连续易位。结果显示每个5'-（TTAGGG）n的模型寡核苷酸‐3'在其展开的MgCl 2溶液中可以产生已解决的电流阻滞。模型寡核苷酸的长度介于6到30个核苷酸之间，与持续时间成正比，在+140 mV处显示出低至0.70-0.13 ms nt -1的转运速度。该方法具有出色的灵敏度和足够的时间分辨率，可深入了解气溶素纳米孔的遗传和表观遗传生物传感方法，使气溶素适用于长链结构化核酸的实际诊断。
更新日期：2018-03-30
中文翻译：
溶血素纳米孔检测二级结构寡核苷酸的一般策略。
气溶素纳米孔被用作对短寡核苷酸（≤10个核苷酸），聚乙二醇（PEG），肽和蛋白质进行单分子分析的灵敏工具。但是，直接分析具有二级结构（例如，G-四链体拓扑）的长寡核苷酸仍然是一个挑战，这阻碍了溶血素纳米孔的进一步实际应用。在这里，提出了一种简单而适用的气溶素纳米孔方法，以实现对结构化寡核苷酸的直接分析，该结构化寡核苷酸通过阳离子调节机制延伸至30个核苷酸长。通过调节电解质溶液中的阳离子类型，结构化的寡核苷酸可展开为线性形式，从而确保连续易位。结果显示每个5'-（TTAGGG）n的模型寡核苷酸‐3'在其展开的MgCl 2溶液中可以产生已解决的电流阻滞。模型寡核苷酸的长度介于6到30个核苷酸之间，与持续时间成正比，在+140 mV处显示出低至0.70-0.13 ms nt -1的转运速度。该方法具有出色的灵敏度和足够的时间分辨率，可深入了解气溶素纳米孔的遗传和表观遗传生物传感方法，使气溶素适用于长链结构化核酸的实际诊断。
京公网安备 11010802027423号