Saha S, Kansal R, Koundal K R
National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi, India.
Indian J Exp Biol. 1996 Oct;34(10):1019-25.
A simple and reliable method was undertaken for the use of polymerase chain reaction in analyzing cDNA clones. Amplification was done of the inserts from positive legumin clones isolated from a cDNA library constructed from developing chickpea cotyledons in the expression vector, gt11. Amplification was made simple by using oligonucleotide primers which allowed convenient sizing, subcloning and sequencing of inserts by di-deoxy chain termination method. This simple method may provide opportunity to isolate large number of agronomically important genes from gene libraries.
采用了一种简单可靠的方法，用于在分析cDNA克隆时使用聚合酶链反应。对从由发育中的鹰嘴豆子叶构建的cDNA文库中分离出的、插入到表达载体gt11中的阳性豆科植物克隆的插入片段进行了扩增。通过使用寡核苷酸引物使扩增变得简单，这些引物便于通过双脱氧链终止法对插入片段进行大小测定、亚克隆和测序。这种简单的方法可能为从基因文库中分离大量具有重要农艺学意义的基因提供机会。
