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通过直接向聚合酶链反应(PCR)反应混合物中添加蔗糖和甲酚红来改进免疫球蛋白重链(IgH)PCR克隆分析。

Modification of IgH PCR clonal analysis by the addition of sucrose and cresol red directly to PCR reaction mixes.

作者信息

Hodges E, Boddy S M, Thomas S, Smith J L

机构信息

Wessex Immunology Service, Southampton University Hospitals NHS Trust, United Kingdom.

出版信息

Mol Pathol. 1997 Jun;50(3):164-6. doi: 10.1136/mp.50.3.164.

Abstract

Diagnostic immunoglobulin (Ig) polymerase chain reaction (PCR) clonality analyses need to be simple, reproducible, and rapid. Sucrose and cresol red (gel loading buffer reagents) were added to a routine IgH PCR reaction mix to obviate the need for adding gel loading buffer separately after PCR amplification. Not only did this decrease the time spent after PCR analysis but also gave similar or enhanced IgH PCR product intensity compared with normal IgH PCR conditions on polyacrylamide gel electrophoresis. This procedure was easily adapted to routine PCR analysis without the need for further manipulations or optimisation of the PCR reaction mix, and it increased the reproducibility and specificity of the IgH PCR products.

摘要

诊断性免疫球蛋白(Ig)聚合酶链反应(PCR)克隆性分析需要具备简单、可重复和快速的特点。将蔗糖和甲酚红(凝胶加样缓冲液试剂)添加到常规的IgH PCR反应混合物中,从而无需在PCR扩增后单独添加凝胶加样缓冲液。这不仅减少了PCR分析后的时间,而且在聚丙烯酰胺凝胶电泳上与正常IgH PCR条件相比,IgH PCR产物强度相似或有所增强。该方法易于应用于常规PCR分析,无需对PCR反应混合物进行进一步操作或优化,并且提高了IgH PCR产物的可重复性和特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cf8/379613/2f8b7e737c6f/molpath00003-0055-a.jpg

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