摘要
用常规RT PCR法 ,以 1组人抗体重链和轻链简并引物 ,直接从 5名丙型肝炎患者外周血混合淋巴细胞中扩增出抗体重链Fd基因和κ轻链基因。结果提示 ,用Trizol试剂提取细胞总RNA ,以Olig(dT)引导合成cDNA第一链 ,再经PCR扩增的方法 ,是值得优先采用的方法。细胞总RNA的提取不必追求高纯度 ,少量外周血即足以满足构建抗体库所需 ,简并引物可获得良好的扩增效果 。
Employing routine RT?PCR,the genes of human immunoglobulin Fd and κ light chains were amplified from hepatitis C patients peripheral blood lymphocytes by sets of oligonucleotide primers. These results suggest that by using Trizol reagant isolated the total RNA of the cells, using Olig(dT) primer reverse transcription synthesized the cDNA first?strand were satisfactory. The high purification of the total RNA of the cells is not necessary to RT?PCR.
出处
《南京铁道医学院学报》
2000年第1期17-20,共4页
Journal of Nanjing Railway Medical College
基金
江苏省科技发展基金资助项目!(BJ97070)
关键词
抗体基因
外周血淋巴细胞
丙型肝炎
PCR
polymerase chain reaction
antibody genes
peripheral blood lymphocytes
