通过在原始方案中增加一个加热步骤，显著提高了基因表达序列分析（SAGE）的克隆效率。
Substantially enhanced cloning efficiency of SAGE (Serial Analysis of Gene Expression) by adding a heating step to the original protocol.
作者信息
Kenzelmann M, Mühlemann K
机构信息
Institute of Medical Microbiology, University of Bern, Friedbühlstrasse 51, 3010 Bern, Switzerland.
出版信息
Nucleic Acids Res. 1999 Feb 1;27(3):917-8. doi: 10.1093/nar/27.3.917.
Abstract
The efficiency of the original SAGE (Serial Analysis of Gene Expression) protocol was limited by a small average size of cloned concatemers. We describe a modification of the technique that overcomes this problem. Ligation of ditags yields concatemers of various sizes. Small concatemers may aggregate and migrate with large ones during gel electrophoresis. A heating step introduced before gel electrophoresis breaks such contaminating aggregates. This modification yields cloned concatemers with an average size of 67 tags as compared to 22 tags by the original protocol. It enhances the length of cloned concatemers substantially and reduces the costs of SAGE.
摘要
原始的基因表达序列分析（SAGE）技术的效率受到克隆串联体平均长度较短的限制。我们描述了一种技术改进方法，可克服这一问题。双标签连接产生各种大小的串联体。小的串联体在凝胶电泳过程中可能会与大的串联体聚集并一起迁移。在凝胶电泳前引入一个加热步骤可打破这种污染性聚集。与原始技术产生平均长度为22个标签的克隆串联体相比，这种改进方法产生的克隆串联体平均长度为67个标签。它显著提高了克隆串联体的长度，并降低了SAGE技术的成本。
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