相关实验视频

Updated: Jul 18, 2025

Novel Sequence Discovery by Subtractive Genomics

Novel Sequence Discovery by Subtractive Genomics

Published on: January 25, 2019

基于DNA或蛋白质序列比较的PCR退化原始物的优化设计

Maria Jorge Campos¹, Alejandro Gallardo², Alberto Quesada²

¹MARE-Marine and Environmental Sciences Centre & ARNET-Aquatic Research Network Associated Laboratory, ESTM, Polytechnic of Leiria, Peniche, Portugal. mcampos@ipleiria.pt.

Methods in molecular biology (Clifton, N.J.)

|August 22, 2023

概括

本研究详细介绍了聚合酶连锁反应 (PCR) 的合理原料设计,以提高效率和特异性. 优化的退行原始剂可以改善基因识别和基因家族内的变异检测.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物信息学是一种生物信息学.

背景情况:

聚合酶链反应 (PCR) 中的退化原始物对于识别未知的蛋白质编码序列和检测基因家族内的遗传变异至关重要.
原料优化,包括战略不匹配引入和位置平衡,提高PCR效率和针对目标序列放大的特异性.

研究的目的:

为三个不同的PCR应用提供合理退化原料设计的详细示例.
为了说明提高复杂遗传分析中的PCR效率和特异性的方法.

主要方法:

退化原料原料的合理设计,在3'-end.具有特定的决定因素.
对于放大相关基因家族成员,3'端的完全退化和5'端的减少退化的应用.
使用5'-end完全退化的原始原料来优化密切相关的正统DNA序列的等位基因歧视.
针对大型基因家族内的特定基因组,以提高PCR效率,使用多重/退化PCR.

主要成果:

通过向退化证明了PCR效率的提高,通过向退化放大相关基因家族成员.
实现了对密切相关的正统DNA序列的等位基因歧视的优化特异性.
展示了不同基因家族内的特定基因组的PCR效率的提高,使多重/退化PCR成为可能.

结论:

退化原料原料的合理设计是提高PCR效率和特异性的强大策略.
优化的原料设计有助于精确的基因识别,变体检测和遗传学分析.
提出的方法为复杂的分子生物学和遗传研究挑战提供了多功能解决方案.

关键词:

在3′末端的3′末端.在5′末端的5′末端.退化原料原料是为了使它们退化.这是一个PCRPCR.在PCR效率方面,PCR效率是非常高的.对于PCR的特异性序列对齐方式序列对齐方式

更多相关视频

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Published on: May 22, 2012

Rapid Characterization of Genetic Parts with Cell-Free Systems

Rapid Characterization of Genetic Parts with Cell-Free Systems

Published on: August 30, 2021

相关概念视频

PCR

PCR

DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

RACE - Rapid Amplification of cDNA Ends

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...

Sanger Sequencing

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...

Evolutionary Relationships through Genome Comparisons

Evolutionary Relationships through Genome Comparisons

Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...