Launer G A, Lukyanov K A, Tarabykin V S, Lukyanov S A
M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Mol Gen Mikrobiol Virusol. 1994 Nov-Dec(6):38-41.
We describe a novel simple PCR-based technique for construction of cDNA libraries starting from the small samples of cells or tissues. This technique is based on the insertion of inverted terminal repeats (ITR) into amplified cDNA which causes a part of molecules to generate "pan"-type structures at each cycle of PCR amplification. This allows to avoid generation of the primer dimmer and makes possible the regulation of an average length of amplified sequences varying in concentration of primers.
我们描述了一种基于聚合酶链式反应(PCR)的新颖且简单的技术,用于从细胞或组织的小样本构建互补DNA(cDNA)文库。该技术基于将反向末端重复序列(ITR)插入到扩增的cDNA中,这使得部分分子在PCR扩增的每个循环中产生“泛”型结构。这有助于避免引物二聚体的产生,并能够通过调节引物浓度来调控扩增序列的平均长度。