PCR-based immortalization and screening of hierarchical pools of cDNAs.
PCR-based immortalization and screening of hierarchical pools of cDNAs.
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基于 PCR 的永生化和 cDNA 分层库的筛选。
DOI：
10.1093/nar/22.22.4806
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发表时间：
1994
影响因子：
14.9
通讯作者：
Schlessinger,D
中科院分区：
文献类型：
--
作者：
D'Esposito,M;Mazzarella,R;Pengue,G;Jones,C;D'Urso,M;Schlessinger,D
从至少60 bp的序列开始，基于pcr的筛选已经发展到从文库中恢复DNA，而不需要杂交或大量的DNA提取步骤。该方法保持了不确定的可用性，即使是稀缺的cDNA文库，并提供了mRNA物种相对丰度的估计。cDNA克隆的分离可以在不到一周的时间内完成。分离的cdna与表达序列片段和基因组序列预测的外显子同源。
Starting from sequences of at least 60 bp, PCR-based screening has been developed to recover cDNAs from libraries without the necessity for hybridization or extensive DNA extraction steps. The method maintains the indefinite availability of even scarce cDNA libraries and provides an estimate of the relative abundance of the mRNA species. Isolation of a cDNA clone can be done in less than a week. cDNAs were isolated that were cognate for fragments of expressed sequences and for an exon predicted from genomic sequence.