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PCR/qPCR耗材選擇——顏色篇

時間:2022/3/18閱讀:546
分享:

PCR/板有多種顏色,如透明色、白色、黃色、藍色、紅色……普通的PCR過程中,管/板的顏色并不會影響PCR的結(jié)果,但是在qPCR中,管/板的顏色對于結(jié)果是有很大的影響的。

應(yīng)用于qPCR的耗材,一般是白色或者磨砂的塑料耗材。qPCRDNA擴增反應(yīng)中,以熒光化學(xué)物質(zhì)測每次PCR循環(huán)后產(chǎn)物總量,因此與透明塑料管/板相比,白色或磨砂的管/板在應(yīng)對熒光時表現(xiàn)更為靈敏,并且結(jié)果更加準(zhǔn)確。這是因為白色的耗材可以阻止熒光折射出管,達到熒光折射的最小化,將更多的熒光信號反射回檢測器,從而增加信噪比,表現(xiàn)為更高的靈敏度;另外,白色管壁也可阻止熒光信號被PCR儀的加熱模塊吸收或進一步反射,從而可以降低重復(fù)實驗中的誤差,提高準(zhǔn)確性。除此之外,在進行qPCR時需使用超透明的管蓋或光學(xué)密封膜來密封PCR/板,保證熒光不受干擾;還需要參考qPCR的耗材要求,選擇最合適的耗材,達到*的實驗效果。


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