之间的区别PCR引物and Sequencing Primers

Key Difference – PCR Primers vs测序Primers

随着分子生物学领域的最新发展，开发了不同的遗传技术，使该受试者不同途径的研究过程变得容易，准确。PCRand other sequencing procedures are two important such techniques. They use different subcomponents.Primersare considered as the major sub-component common to both PCR and Sequencing techniques.PCR primers are used for amplification of a particularDNA序列whilst s方程引物用于对DNA片段进行测序的背景，以揭示其特定顺序核苷酸序列。这是关键区别between PCR primers and sequencing primers.

CONTENTS

1。概述和关键差异
2。What are PCR Primers
3。What are Sequencing Primers
4。Similarities Between PCR Primers and Sequencing Primers
5.Side by Side Comparison – PCR Primers vs Sequencing Primers in Tabular Form
6.Summary

什么是PCR底漆？

Polymerase Chain Reaction (PCR) is a genetic technique that is utilized in the field of molecular biology in order to amplify a single or few copies of a particular DNA segment and to obtain many millions of identical copies. In a PCR reaction, different components are used including primers. Primers are short DNA strands with a nucleotide length of 18-25 making them compatible with the start and the end region of the DNA fragments to be amplified. Primers can be a forward primer and reverse primer. These primers bind to the DNA fragment at the specific points where it makesDNA聚合酶to bind to the specific primer at the location and initiate the synthesis of the new DNA strand.

底漆的选择是PCR过程的重要方面。底漆长度的选择很重要。理想的长度为18-25个核苷酸。如果长度太短或太长，则引物不会与DNA序列结合以准确放大。长度太短的引物会导致在DNA序列的不同位置进行非特异性引物退火。

这鸟嘌呤andCytosine(GC) content in a good primer should be in the range of 40-60. The primer annealing temperature and melting temperature are vital factors during PCR. The melting temperature should be calculated accurately, and the primer annealing temperature should be 5⁰C小于熔化温度。熔化温度应为60°C和75°C。温度过高或太低会导致活性DNA聚合酶活性较低。

什么是测序引物？

测序引物用于测序DNA片段的上下文，目的是揭示其特定的身份。为了获得良好的测序结果，高质量的引物和模板很重要。Thus, when primers are selected, they should be unique to a particular region where we desire to sequence.它也应该具有正确的方向，即序列通常从引物的3'至5'端产生。该序列应缺乏不良的自脑化，例如发夹回路的形成。它不应连续形成鸟嘌呤碱。

这melting temperature (Tm) of the primer must be suitable for the conditions of the sequencing. Therefore, it should lie between 52^oC和74^oC. Preparation ofoligonucleotides应纯化用作底漆以获得所需的序列的全长。如果寡核苷酸含有杂质，则底漆序列信号传导将从不同的启动位点叠加，并且还将减少碱细胞的数量。

oligonuc的引物熔化温度(Tm)leotide determines, how strong the complementary DNA strands are hybridized with each other. Tm can be considered as a thermodynamic calculation where it is dependent on both DNA sequences and several conditions such as salt concentration. The Tm is important during PCR where a variant called the cycle sequencing is used to produce a group of dideoxynucleotide-terminated fragments. Here, the primer that is sequenced will initially be annealed alternatively, then extended and lastly denatured for amplification. Therefore, the Tm value should be in between 52^oC和74^oC.根据选择，可以从DNA/RNA合成实验室获得合成的寡核苷酸。用于DNA测序的小规模合成通常为50 nmol。同样，最重要的是，应将用于测序的底漆纯化为没有杂质，以防止质量降低。

What are the Similarities Between PCR Primers and Sequencing Primers?

• PCR引物和测序引物都是引物，用于靶向DNA序列的放大过程中。
• PCR引物和测序引物均由核苷酸组成。
• PCR引物和测序引物都是短的低聚物。

What is the Difference Between PCR Primers and Sequencing Primers?

PCR引物与测序Primers
PCR primers are short DNA strands with a nucleotide sequence length of 18-25 making them compatible with the start and the end region of the DNA fragments that are to be amplified.	测序引物是在测序DNA片段的背景下使用的短寡聚物，目的是揭示其特定的身份。
Function
PCR primers are used for amplification of a particular DNA sequence.	测序引物用于测序DNA片段的上下文，目的是揭示其特定的身份。
Number of primers Needed
Two primers; one forward primer and one reverse primer are used as PCR primers.	Need only one primer as sequencing primer.

概括 -PCR引物与测序Primers

测序引物用于测序DNA片段的上下文，目的是揭示其特定的身份。One sequencing primer will be enough to run the process. To obtain good sequencing results, high quality primers and templates are important. Thus, when primers are selected, they should be unique to a particular region where we desire to sequence. PCR Primers are short DNA strands with a nucleotide length of 18-25 which is compatible with the start and the end region of the DNA fragments that are to be amplified. PCR primers can be a forward primer and reverse primer. The Guanine and Cytosine (GC) content in a good primer should be in the range of 40-60. The primer annealing temperature and melting temperature are vital aspects during PCR. This is the difference between PCR primers and Sequencing primers.

参考：

1。“Polymerase chain reaction (PCR).” Khan Academy.在这里可用
2。“Sequencing primers and primer design.” Sequencing primers and primer design | University Core DNA Services | University of Calgary.在这里可用

图片提供：

1.’Primers revcomp’sepy Zephyris - 自己的工作，（CC BY-SA 3.0）通过下议院维基梅迪亚
2。’DNA Sequencin 3 labeling methods’By Abizar (original uploader) at English Wikipedia – Transferred by Gustavocarra., (Public Domain) via下议院维基梅迪亚