之间的区别Nick Translation and End Filling

这关键区别在尼克翻译和结束填充之间nick translation is a process that creates labelled DNA探针for various hybridization reactions while end filling is a technique that creates blunted fragments by adding核苷酸进入单链悬垂。

尼克翻译和末端填充是分子生物学中使用的两种技术。Nick Translation用于标记杂交探针，以检测特定的核苷酸序列。末端填充用于制作带有单链悬垂的粘性末端的钝片段。两种技术都非常重要，并且它们在分子研究实验室中通常进行。

1。概述和关键差异
2。什么是尼克翻译
3。什么结局填充
4。尼克翻译与结束填充之间的相似之处
5。并排比较 - 划痕与最终填充表格形式
6。Summary

什么是尼克翻译？

Nick Translation是一种重要的技术，用于制备各种分子生物学技术（例如吸毒，，，，原位杂交，荧光原位杂交等。这是一个in vitroDNA标记方法。DNA探针用于鉴定特定的DNA或RNA序列。借助标记的探针，可以从核酸的复杂混合物中标记或可视化特定的片段。因此，使用各种技术制备标记的探针。Nick Translation就是一种这样的方法，它在DNase 1和DNA聚合酶1酶的帮助下产生标记的探针。

图01：尼克翻译

Nick translation process starts with DNase 1 enzyme activity. DNase 1 introduces nicks to the phosphate backbone of double-stranded DNA by cleaving phosphodiester bonds between nucleotides. Once the nick is created, free 3′ OH group of the nucleotide will be produced, and DNA polymerase 1 enzyme will act on it. 5′ to 3′ exonuclease activity of DNA polymerase 1 removes nucleotides from the nick towards the 3′ direction of the DNA strand.

同时，DNA聚合酶1酶的聚合酶活性起作用，并添加核苷酸以替代去除的核苷酸。如果标记核苷酸，则将通过标记的核苷酸进行替换，并将其标记为识别DNA。最后，这种新合成的标记DNA可以用作各种杂交反应中的探针。

什么结局填充?

灌装技术用于分子biolo结束gy to make blunted fragments. Restriction digestion produces fragments with overhangs. These fragments may not be compatible to ligate into plasmid vectors. Vectors are often blunted to allow non-compatible ends to be joined. Therefore, fragments with overhangs can be blunted by adding the nucleotides to the complementary strand using the overhang as the template for polymerization. This process is known as end filling.

DNA聚合酶，例如DNA聚合酶I和T4 DNA聚合酶的Klenow片段可催化末端填充。它们添加核苷酸以填充（5'→3'）并咀嚼后（3'→5'）。一旦粘性末端填充，它们就会变得钝，并准备将其结合到矢量中。

Furthermore, end filling can be used to label DNA molecules. However, it can only be used to label DNA molecules with sticky ends. In comparison to nick translation, end filling is a gentler method which rarely causes DNA breaks.

What are the Similarities Between Nick Translation and End Filling?

尼克翻译和末端填充都是分子生物学技术。
它们可用于标记探针。
Both arein vitro

What is the Difference Between Nick Translation and End Filling?

Nick Translation是一种创建标记为杂交探针的技术。相比之下，末端填充是一种创造钝器片段的技术，用于将其连接到向量中。因此，这是划痕翻译和结束填充之间的关键区别。此外，划痕翻译需要使用5'TO 3'外切核酸酶活性，而末端填充不需要5'TO 3'核酸外切酶活性。

这below infographic tabulates the differences between nick translation and end filling in more detail.

摘要 - 尼克翻译与最终填充

Nick translation is a method that incorporates radiolabeled nucleotides into DNA. It synthesizes labelled probes based on the activities of DNase 1 and大肠杆菌DNA聚合酶1酶。另一方面，最终填充是一种产生钝片段的技术。当碎片具有粘性末端（单链悬垂）时，有必要使它们钝化的末端以将其结合到矢量中。末端填充增加了兼容的核苷酸并创建钝的末端片段，以使其与连接兼容。因此，这总结了划痕翻译和结束填充之间的差异。