利用干血斑进行突变分析的快速、廉价的基因分型方法在突变小鼠模型中的应用：最新进展。

A rapid and inexpensive genotyping method using dried blood spots for mutational analysis in a mutant mouse model: an update.

机构信息

Department of Biology, University of Naples "Federico II", Via Cinthia, 80126, Naples, Italy.

Pathology Unit, Department of Mental and Physical Health and Preventive Medicine, University of the studies of Campania "L. Vanvitelli", via Luciano Armanni 5, 80138, Naples, Italy.

出版信息

Mol Biol Rep. 2022 Sep;49(9):9071-9077. doi: 10.1007/s11033-022-07649-x. Epub 2022 Jun 22.

DOI:10.1007/s11033-022-07649-x

PMID:35733059

Abstract

BACKGROUND

Dried blood spot (DBS) testing is a well-known method of bio-sampling by which blood samples are blotted and dried on filter paper. The dried samples can then be analyzed by several techniques such as DNA amplification and HPLC. We have developed a non-invasive sampling followed by an alternative protocol for genomic DNA extraction from a drop of blood adsorbed on paper support. This protocol consists of two separate steps: (1) organic DNA extraction from the DBS, followed by (2) DNA amplification by polymerase chain reaction (PCR). The PCR-restriction fragment length polymorphism (PCR-RFLP) is an advantageous and simple approach to detect single nucleotide polymorphisms (SNPs).

RESULTS

We have evaluated the efficiency of our method for the extraction of genomic DNA from DBS by testing its performance in genotyping mouse models of obesity and herein discuss the specificity and feasibility of this novel procedure.

CONCLUSIONS

Our protocol is easy to perform, fast and inexpensive and allows the isolation of pure DNA from a tiny amount of sample.

摘要

背景

干血斑（DBS）检测是一种众所周知的生物采样方法，通过该方法可以将血液样本点在滤纸上并使其干燥。干燥的样本随后可以通过几种技术进行分析，例如 DNA 扩增和 HPLC。我们开发了一种非侵入性采样方法，随后开发了一种从吸附在纸支持物上的血液滴中提取基因组 DNA 的替代方案。该方案包括两个单独的步骤：（1）从 DBS 中提取有机 DNA，然后（2）通过聚合酶链反应（PCR）进行 DNA 扩增。聚合酶链反应-限制性片段长度多态性（PCR-RFLP）是一种有利且简单的方法，可用于检测单核苷酸多态性（SNP）。