基于短串联重复序列分析的等温扩增DNA质量控制

Quality Control of Isothermal Amplified DNA Based on Short Tandem Repeat Analysis.

作者信息

Kroneis Thomas, El-Heliebi Amin

机构信息

Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21, 8010, Graz, Austria.

Sahlgrenska Cancer Center, University of Gothenburg, Gothenburg, Sweden.

出版信息

Methods Mol Biol. 2015;1347:129-40. doi: 10.1007/978-1-4939-2990-0_10.

DOI:10.1007/978-1-4939-2990-0_10

PMID:26374315

Abstract

This protocol describes the use of a 16plex PCR for the purpose assessing DNA quality after isothermal whole genome amplification (WGA). In short, DNA products, generated by amplification multiple displacement amplification, are forwarded to PCR targeting 15 short tandem repeats (STR) as well as amelogenin generating up to 32 different PCR products. After amplification, the PCR products are separated via capillary electrophoresis and analyzed based on the obtained DNA profiles. Isothermal WGA products of good DNA quality will result in DNA profiles with efficiencies of >90 % of the full DNA profile.

摘要

本方案描述了使用16重PCR来评估等温全基因组扩增（WGA）后DNA质量的方法。简而言之，通过多重置换扩增产生的DNA产物被用于靶向15个短串联重复序列（STR）以及牙釉蛋白的PCR，从而产生多达32种不同的PCR产物。扩增后，通过毛细管电泳分离PCR产物，并根据获得的DNA图谱进行分析。DNA质量良好的等温WGA产物将产生效率>90%的完整DNA图谱的DNA图谱。