λgt10 cDNA文库表达序列标签测定时模板处理方式

Processing Methods of Expressed Sequence Tags Sequencing Template for λgt10 cDNA Library

表达序列标签是揭示基因组容量的有效方法。在构建 c DNA文库的基础上 ,对以 λgt1 0为载体进行表达序列标签测定时模板的处理方法进行了探讨。结果表明 ,以λ噬菌体 DNA为模板直接测序比 PCR产物经回收后为模板进行测序 ,其目标克隆的平均插入片段长度要长 ,但反应成功率低。以 λ噬菌体浸提液为模板进行 PCR并在产物回收后进行测序反应 ,可能是以

The generation of expressed sequence tags(ESTs) has been proven to be a rapid and efficient approach by which to identify expressed genes and characterize the information content of genome.On the base of cDNA library construction,the processing methods for λgt10 vector template were selected for ESTs sequencing.The results showed that the insert fragment length of target clones as λ phage DNA template directly sequencing was longer than that of PCR products after recycling,but a lower succeeding ratio for sequencing reaction.The sequencing template of PCR products with λ phage elution may be a best EST sequencing method for cDNA library constructed with λ phage vector.