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基因组尾部DNA的狭缝印迹法。

Slot blotting of genomic tail DNA.

作者信息

Murphy D

机构信息

Neuropeptide Laboratory, Institute of Molecular and Cell Biology, National University of Singapore, Republic of Singapore.

出版信息

Methods Mol Biol. 1993;18:313-5. doi: 10.1385/0-89603-245-0:313.

Abstract

Slot or dot blotting is a technique whereby nucleic acids can be applied to a solid matrix, unfractionated, using a vacuum manifold. Slot blotting is the quickest, easiest, and, apart from polymerase chain reaction, probably the most sensitive assay of transgenic animal genotype (1). However, it can only be applied to animals that bear a transgene that has hybridizable segments with little or no homology to the host genomic DNA-for example, a hybrid gene with a viral or procaryotic reporter element, or a gene from another species with sufficient sequence divergence to allow the transgene to be distinguished from the host gene. Genomic slot blotting should not be used for the initial identification of a transgenic founder animal. For this, Southern blotting is preferable as it can give more information about the number of integration sites and the presence of transgene rearrangements, deletions, and so forth. However, genomic slot blotting is useful for the rapid screening of subsequent generations of transgenics when detailed information about the structure of the transgene is not needed.

摘要

狭缝或斑点印迹法是一种可利用真空歧管将未分级的核酸应用于固体基质的技术。狭缝印迹法是转基因动物基因型检测中除聚合酶链反应外最快、最简单且可能最灵敏的检测方法(1)。然而,它仅适用于携带与宿主基因组DNA几乎没有同源性或同源性很低的可杂交片段的转基因动物,例如带有病毒或原核报告元件的杂交基因,或来自另一个物种且具有足够序列差异从而使转基因能够与宿主基因区分开来的基因。基因组狭缝印迹法不应用于转基因奠基动物的初步鉴定。为此,Southern印迹法更可取,因为它能提供更多关于整合位点数量以及转基因重排、缺失等情况的信息。然而,当不需要有关转基因结构的详细信息时,基因组狭缝印迹法对于快速筛选转基因后代很有用。

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