Sat, 26 Jul 2008 08:07:10 BST CiteULike: vrich's library [222 articles] http://www.citeulike.org/user/vrich/article/556231 CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/vrich/article/556231 <i>Appl Environ Microbiol, Vol. 71, No. 12. (December 2005), pp. 8966-8969.</i><br /><br />The contribution of PCR artifacts to 16S rRNA gene sequence diversity from a complex bacterioplankton sample was estimated. Taq DNA polymerase errors were found to be the dominant sequence artifact but could be constrained by clustering the sequences into 99% sequence similarity groups. Other artifacts (chimeras and heteroduplex molecules) were significantly reduced by employing modified amplification protocols. Surprisingly, no skew in sequence types was detected in the two libraries constructed from PCR products amplified for different numbers of cycles. Recommendations for modification of amplification protocols and for reporting diversity estimates at 99% sequence similarity as a standard are given. PCR-induced sequence artifacts and bias: insights from comparison of two 16S rRNA clone libraries constructed from the same sample. SG Acinas R Sarma-Rupavtarm V Klepac-Ceraj MF Polz doi:10.1128/AEM.71.12.8966-8969.2005 Appl Environ Microbiol, Vol. 71, No. 12. (December 2005), pp. 8966-8969. 2006-03-17T19:55:57-00:00 2005 Appl Environ Microbiol 0099-2240 71 12 8966 8969 pcr pcr_bias