交叉探头 - - 通过模板增强的杂交工艺对核酸进行序列特定检测
Shizuka Nakayama1, Lei Yan, Herman O Sintim
1Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Journal of the American Chemical Society
|September 2, 2008
概括
交叉探头技术使得在恒温下放大核酸检测成为可能. 将第二个维度添加到探针中,可以让廉价的酶检测单核酸多态 (SNP).
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 遗传学 是一个遗传学.
背景情况:
- 核酸检测技术对于分子诊断至关重要.
- 同热放大方法比传统的PCR具有优势.
- 单核酸多态 (SNP) 是重要的遗传标记.
研究的目的:
- 为了引入一种新的连接探头核酸检测技术.
- 为了在同热条件下实现放大分析器传感.
- 为了利用随时可用的DNA处理酶进行SNP检测.
主要方法:
- 交叉探针核酸检测技术的开发.
- 将第二个维度纳入检测探头.
- 限制性内核酶用于SNP识别的应用.
主要成果:
- 在同热条件下对分析物的放大感应.
- 成功地利用了商业上可用的DNA处理酶.
- 使用新型探针系统实现了单核酸多态 (SNP) 的检测.
结论:
- 交叉探头技术为同热核酸检测提供了一种有效的方法.
- 限制性内核酶的使用为SNP检测提供了一种具有成本效益的方法.
- 这项技术在基因分析和诊断方面具有潜在的应用.
相关概念视频
Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
In-situ Hybridization
In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Southern Blot
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
Maxam-Gilbert Sequencing
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.


