摘要
目的 :通过 PCR扩增从基因组中获得长达 1.42 kb人干细胞因子基因 (SCF) 5’旁侧调控序列 ,为后续表达调控研究做准备。方法 :对常规 PCR缓冲液调节 p H值至 9.0 ,Taq DNA聚合酶中加入适量的具有校正功能的 Tli DNA聚合酶 ,退火温度升至 6 8℃ ,加大模板量 ,增加模板预变性时间 ,进行 2 5个循环。结果 :经过改变条件 ,成功地获得了所需的 1.42 kb的 SCF调控序列片段。结论 :在一般的 PCR体系中添加适量的具有 3’→ 5’外切核酸酶活性和校正功能的Tli DNA聚合酶扩增稍长
Objective [WT5”BZ]To prepare for the following regulation and expression study of the SCF gene,a 1.42 kb 5’ flanking regulation sequence was amplified.[WT5”HZ]Methods:[WT5”BZ]Normal polymerase chain reaction (PCR) buffer pH was adjusted to 9.0,some Tli DNA polymerases which has proofreading function were added,annealing temperature rise to 68℃,the amount of the genomic DNA template increased,then run 25 PCR cycles. [WT5”HZ]Results:[WT5”BZ]SCF 5’ flanking regulation sequence was successfully amplified.[WT5”HZ]Conclusion:[WT5”BZ]It is a economical and effective way to amplify longer DNA fragment after some enzymes which has 3’→5’ exonuclease activity and proofreading function were added to the normal PCR system. [WT5”HZ]
出处
《广东医学院学报》
2000年第2期119-121,共3页
Journal of Guangdong Medical College
基金
国家自然科学基金!项目资助 (NO.3970 0 0 5 0
