PrinciplesofPCR.docVIP

上海海洋大学 食品学院 08制药一班 OH翻译版 I6 POLYMERASE CHAIN REACTION PCR 【聚合酶联反应】 Key Notes Principles of PCR【聚合酶联反应的原理】 ?The polymerase chain reaction PCR allows an extremely large number of copies to be synthesized of any given DNA sequence provided that two oligoonucleotide primers are available that hybridize to the flanking sequences on the complementary DNA strands. 【聚合酶联反应指一种在体外快速扩增特定基因或DNA序列的方法】The reaction requires the target DNA, the two primers, all four deoxyribonucleoside triphosphates and a thermostable DNA polymerase such as Taq DNA polymerase. 【这种反应需要目标DNA，两个引物，四个脱氧和干三磷酸和一个耐热DNA聚合酶例如噬热水生菌DNA聚合酶】A PCR cycle consists of three steps; denaturation, primer annealing and elongation.【一个PCR循环反应包括3个步骤：模板变性】 This cycle is repeated for a set number of times depending on the degree of amplification required.【这个循环将重复一定的次数，而重复的次序是取决于需要扩增的程度】 Applications of PCR【PCR的程度】 ?PCR has made a huge impact in molecular biology, with many applications in areas such as cloning, sequencing, the creation of specific mutations, medical diagnosis and forensic medicine.【PCR技术很大程度上影响了分子生物学的发展，在这个领域上有很大的应用例如克隆，测序，创造特定的基因突变，医疗诊断和法医学】 Related topics?? DNA structure F1 ? 【DNA 结构】 Nucleic acid hybridization I2 ? 【核酸杂交】 DNA replication in prokaryotes F3 ? 【在原核生物中的DNA复制】 DNA cloning I3 ?【DNA克隆】 Restriction enzymes I1 【限制性内切酶】 DNA sequencing I5 ? 【DNA测序】 Principles of PCR【PCR的原理】 PCR polymerase chain reaction is an extremely simple yet immensely powerful technique. 【PCR 聚合酶联反应 一种操作简单，应用强大的技术】It allows enormous amplification of any specific sequence of DNA provided that short sequences either side of it are known. The technique is shown in Fig.15. 【PCR满足任何特定序列的DNA哪怕这条DNA只有一端是一直的】A PCR reaction contains the target double-stranded DNA, two primers that hybridize to flanking sequences on opposing strands of the target, all four deoxyribonucleoside triphosphates and a DNA polymerase. 【PCR反应包括目标双链DNA，用于杂交目标链的反向链开始杂交的两个引物，四种脱氧核苷酸三磷酸和一个DNA聚合酶】Because, as we shall see, the reaction periodically g