High-throughput methods for measuring DNA thermodynamics

被引：19

作者：

Bae, Jin H. ^{[1
]}

Fang, John Z. ^{[1
]}

Zhang, David Yu ^{[1
,2
]}

机构：

[1] Rice Univ, Dept Bioengn, Houston, TX 77005 USA

[2] Rice Univ, Syst Synthet & Phys Biol, Houston, TX 77005 USA

来源：

NUCLEIC ACIDS RESEARCH | 2020年 / 48卷 / 15期

关键词：

DAMAGE; STABILITY;

D O I：

10.1093/nar/gkaa521

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Understanding the thermodynamics of DNA motifs is important for prediction and design of probes and primers, but melt curve analyses are low-throughput and produce inaccurate results for motifs such as bulges and mismatches. Here, we developed a new, accurate and high-throughput method for measuring DNA motif thermodynamics called TEEM (Toehold Exchange Energy Measurement). It is a refined framework of comparing two toehold exchange reactions, which are competitive strand displacement between oligonucleotides. In a single experiment, TEEM can measure over 1000 Delta G degrees. values with standard error of roughly 0.05 kcal/mol.

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页数：9

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