摘要:1.IntroductionPolymerase chain reaction (PCR) has rapidly become one of the most widely used techniques in molecular biology and for good reason: it is a rapid, inexpensive and simple means of produci[阅读全文:]
摘要: PCR反应的关键因素主要有引物的选择与设计，酶的质量。模板的制备，在前二者都稳定可行的情况下，PCR模板的制备尤为重要。模板处理方法的选择及操作人员的基 本技能，决定分离模板核酸(DNA或RNA)的质和量及PCR的成败。而提[阅读全文:]
摘要:1.protocol(1)collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH(2)put in boiling H2O for 30 sec (optimum may need to be determined for each type [阅读全文:]
摘要:Designing PCR primers1.length of individual primers between 18-24 bases. Longer primers (30-35 bp) seem to work in more similar cycling conditions compared with shorter primers, and can make multiplex[阅读全文:]
摘要:1.DMSO and glycerolRecommended by a number of authors to improve amplification efficiency and specificity of PCR. However,in the multiplex reaction these enhancers gave conflicting results.For example[阅读全文:]
摘要:热启动PCR热启动PCR是除了好的引物设计之外，提高PCR特异性最重要的方法之一。尽管Taq DNA聚合酶的最佳延伸温度在72℃，聚合酶在室温仍然有活性。因此，在进行PCR反应配制过程中，以及在热循环刚开始，保温温度低于退火温度[阅读全文:]
摘要:1、引物设计由美国Perkin-elmer公司合成，引物末端标记有生物素（1）扩增体系10×PCR buffer5μlPCR引物10μl4×dNTP10μlTaq polymerase0.25μlDNA模板10μlD、W14.75μlTotal50μl（2） 扩增程序[阅读全文:]
摘要:多聚酶链式反应(polymerase chain reaction)简称PCR技术，是80年代中期发展起来的一种体外扩增特异DNA片段的技术。此法操作简便， 可在短时间内在试管中获得数百万个特异的目的DNA序列的拷贝，PCR技术虽然问世仅数年时间[阅读全文:]