PCR inhibition by reverse transcriptase leads to an overestimation of amplification efficiency

Content Provider	Semantic Scholar
Author	Suslov, Oleg N.
Copyright Year	2005
Abstract	This study addresses the problem of PCR inhibition by reverse transcriptase. It has been shown that the inhibition occurs mostly when a small amount of RNA is taken for RT reaction, and it is more visible for rarely expressed transcripts. We show here that the inhibition takes place regardless of what amount of template is utilized for RT. The inhibition possesses a global nature, i.e. the amplification of any given transcript may be compromised with different levels of inhibition. The process of inhibition also explains wrongfully derived PCR amplification efficiencies, sometimes more than 100%, when the sequential dilutions of unpurified RT sample are utilized to build the calibration curve. The RT influences PCR not only by inhibiting it. When microgram(s) of RNA are taken for RT reaction, reverse transcriptase may cause overamplification of some transcripts under certain PCR conditions. The possible mechanism of RT influence on PCR is presented, and a purification method is implemented to remove the effects of RT on PCR.
Starting Page	e181
Ending Page	e181
Page Count	1
File Format	PDF HTM / HTML
PubMed reference number	16314311
Volume Number	33
Journal	Nucleic acids research
Alternate Webpage(s)	http://www.gene-quantification.com/suslov-eff-nar-2005.pdf
Alternate Webpage(s)	https://watermark.silverchair.com/gni176.pdf?token=AQECAHi208BE49Ooan9kkhW_Ercy7Dm3ZL_9Cf3qfKAc485ysgAAAj8wggI7BgkqhkiG9w0BBwagggIsMIICKAIBADCCAiEGCSqGSIb3DQEHATAeBglghkgBZQMEAS4wEQQMnpBtM0J5oSupum4EAgEQgIIB8gdmPfDKvaKkDf_ZKbLM17lXJ6O3mpdRHwHNrokJmtCBzJ82VMsQgDAANv1PM06ErOjazDi_pUiLEslhM30bBUKUU4ErtqCfSULsa28MjjWZZWh3w1wwjov6BFU2n-XV4u7ZXg07ItZ1CX2_ERnlTgnxDt5V5GYj9chC_phFNHUdgTgxKF8Tuq1Ro6QTc08R2Sakyy-1PorYkadWgZ-utCTGkhm2p9iWagDPSm997mn2zbze8aDhYaveVvUGXT2R1N9Poa7lxsHfOm5s5ANUsDuef0EjTXdKeOuUBd5Bc8Ngjsmev4jcL7qp657XNQQV_AxjvAREozFyEUvaiPIil_FqTDsLKXCKCokkvocrV8ssJDQKEexDWp4lQGtvOZCC45J66OdsLuJefRUg3APadBVHIwoiySEEpJ_7K07FN_wkH8Lt5NLSv8XnbG_NnuU_CeQmn5hzj2-W4aMTVgnreNl9CdIzw4oG6jUkFmmWfWnnH67UglA8FezxkB3xMrEx4Han0FjvIaLk7CHge2I5mG_t9hETUOPOTKqIIiYc872Si0UiHuZV5k7iSJyPM8mNuEFmn99svPHyoxFiX3oQ7rO-Fg8zgUWqKclQI-GRyXODSO9Kvir7MILwCJP7Bhgk-GAXer1QY5lysRqT4hSKSE5dxQ
Alternate Webpage(s)	http://nar.oxfordjournals.org/content/33/20/e181.full-text-lowres.pdf
Alternate Webpage(s)	http://nar.oxfordjournals.org/content/33/20/e181.full.pdf
Alternate Webpage(s)	http://www.gene-quantification.de/suslov-eff-nar-2005.pdf
Language	English
Access Restriction	Open
Subject Keyword	Clinical Use Template Gene Amplification Technique Inhibition RNA RNA-Directed DNA Polymerase Transcript microgram
Content Type	Text
Resource Type	Article