Okubo K, Hori N, Matoba R, Niiyama T, Matsubara K
Institute for Molecular and Cellular Biology, Osaka University, Japan.
DNA Seq. 1991;2(3):137-44. doi: 10.3109/10425179109039684.
We have developed a method for constructing a library containing the 3' end fragment of cDNA for large-scale sequencing of cDNA clones. The average size of the insert was 270 bp. Cell lysates that carry plasmids having the cDNA insert were subjected to PCR amplification of the cDNA moiety and the products were subjected to sequencing analysis using an autosequencer. With this protocol, sample preparation became a non-limiting step, that allowed us to sequence as many samples as the autosequencer could handle.
我们开发了一种构建文库的方法，该文库包含用于cDNA克隆大规模测序的cDNA 3'端片段。插入片段的平均大小为270 bp。携带具有cDNA插入片段质粒的细胞裂解物进行cDNA部分的PCR扩增，产物使用自动测序仪进行测序分析。通过该方案，样品制备成为一个非限制性步骤，这使我们能够对自动测序仪所能处理的尽可能多的样品进行测序。
