Sartorius赛多利斯VS-AVPQ502Vivapur

RT-PCR and PCR

Thermocycler can reach temperatures of 95 ℃. RT set-up 1. To a sterile 500 µl Eppendorf tube add in the following order: 2. Incubate 65 ûC for 5 min. 3. Cool slowly

PCR和RT-PCR

(A)+RNA。不管起始模板是总RNA还是poly(A)+ RNA，都可以检测到扩增结果（图2）。另外，分离poly(A)+ RNA会导致样品间mRNA丰度的波动变化，从而使信息的检出和定量产生偏差。然而，当分析稀有mRNA时，poly(A)+ RNA会增加检测的灵敏度。 图2. 总RNA和poly(A)+ RNA在RT-PCR中的比较 以5或1μg Hela细胞总RNA（分别为泳道1和2）或500ng和50ng Hela细胞poly(A)+ RNA（分别为泳道

Protocol for competitive RT-PCR

) and the success of this treatment has to be controlled by a PCR without prior RT. It is, however, always recommendable to treat the RNA samples with DNase so that no genomic DNA competes for the PCR components. To make a standard, first a PCR