DNase pretreatment of master mix reagents improves the validity of universal 16S rRNA gene PCR results.
DNase I pretreatment of 16S rRNA gene PCR reagents was tested. The DNase I requirement for the elimination of false-positive results varied between 0.1 and 70 IU per master mix depending on the applied Taq polymerase. PCR sensitivity was mostly maintained when 0.1 IU of DNase I was used.
Heininger
Alexandra
A
Klinik für Anästhesiologie und Transfusionsmedizin, Abteilung für Anästhesiologie, Eberhard-Karls-Universität Tübingen, D-72074 Tübingen, Germany.
Binder
Marlies
M
Ellinger
Andreas
A
Botzenhart
Konrad
K
Unertl
Klaus
K
Döring
Gerd
G
D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005189:False Positive Reactions; D020459:Genes, rRNA; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000378:metabolism
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D019914:Taq Polymerase; D003850:Deoxyribonuclease I