PMID- 7684388
OWN - NLM
STAT- MEDLINE
DCOM- 19930621
LR  - 20190830
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 41
IP  - 2
DP  - 1993 Feb
TI  - Restriction endonuclease digestion eliminates product contamination in reverse 
      transcribed polymerase chain reaction.
PG  - 235-8
AB  - Restriction endonuclease digestion was used to eliminate false-positive signals 
      caused by polymerase chain reaction (PCR) product DNA contamination in a reverse 
      transcribed (RT) PCR for amplifying rubella virus (RV) RNA sequences. A 
      restriction enzyme selected to cut the PCR product DNA between, but not within, 
      the primer binding sites was used to digest reaction mixtures after reverse 
      transcription but before PCR amplification. Because restriction enzymes generally 
      react only with specific double-strand sequences, contaminating DNA was rendered 
      inactive while reverse-transcribed single strand cDNA was amplified. Assays 
      showed that restriction enzyme digestion reduced template activity of product DNA 
      by a factor of 10(7), while leaving sensitivity of the RT-PCR unaffected.
FAU - Dougherty, R M
AU  - Dougherty RM
AD  - Department of Microbiology and Immunology, SUNY Health Science Center, Syracuse 
      13210.
FAU - Phillips, P E
AU  - Phillips PE
FAU - Gibson, S
AU  - Gibson S
FAU - Young, L
AU  - Young L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA, Viral)
RN  - 0 (RNA, Viral)
RN  - EC 2.7.7.49 (RNA-Directed DNA Polymerase)
RN  - EC 3.1.21.4 (CCCGGG-specific type II deoxyribonucleases)
RN  - EC 3.1.21.4 (Deoxyribonucleases, Type II Site-Specific)
SB  - IM
MH  - Animals
MH  - DNA, Viral/analysis
MH  - Deoxyribonucleases, Type II Site-Specific/*metabolism
MH  - *False Positive Reactions
MH  - Polymerase Chain Reaction/*methods
MH  - RNA, Viral/analysis
MH  - RNA-Directed DNA Polymerase/metabolism
MH  - Rubella virus/genetics/*isolation & purification
MH  - Vero Cells
EDAT- 1993/02/01 00:00
MHDA- 1993/02/01 00:01
CRDT- 1993/02/01 00:00
PHST- 1993/02/01 00:00 [pubmed]
PHST- 1993/02/01 00:01 [medline]
PHST- 1993/02/01 00:00 [entrez]
AID - 0166-0934(93)90130-J [pii]
AID - 10.1016/0166-0934(93)90130-j [doi]
PST - ppublish
SO  - J Virol Methods. 1993 Feb;41(2):235-8. doi: 10.1016/0166-0934(93)90130-j.