Bacterial DNA decontamination for reverse transcription polymerase chain reaction (RT-PCR).
For RT-PCR, removal of contaminating genomic DNA in RNA samples using manganese sulfate was more effective than magnesium. DNA contamination was removed in 3 microg of nucleic acid using 10 U of RNase-free DNase I in 10-microl reaction volumes. The digestion procedure was compatible with commercial RNA extraction kits and was suitable for RT-PCR assay.
Wang
Gehua
G
National Laboratory for Enteric Pathogens, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, Manitoba, Canada R3E 3R2.
Barton
Connie
C
Rodgers
Frank G
FG
D002021:Buffers; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D003850:Deoxyribonuclease I / Q000737:chemistry; D019453:Escherichia coli O157 / Q000235:genetics*; D017895:Manganese Compounds / Q000737:chemistry; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013431:Sulfates / Q000737:chemistry
D002021:Buffers; D004269:DNA, Bacterial; D017895:Manganese Compounds; D012329:RNA, Bacterial; D013431:Sulfates; D003850:Deoxyribonuclease I; C039798:manganese sulfate