Table 1. Contamination of commercial RT-PCR reagents with murine leukemia virus (MLV).1 .
Manufacturer | Kit/Reagent | RT origin | Lot no. | pro (%, copies/rxn) | gag | Nested pol (%) | mtDNA 2 (copies/rxn) | IAP 2 (copies/rxn) |
ABI/Ambion | AgPath One Step RT-PCR kit | MoMLV | 1009030 | 7/16 (44, 1–10) | 0/16 | 6/16 (38) | 0/16 | 0/16 |
AgPath One Step RT-PCR kit | MoMLV | 909020 | 1/16 (6, 1–10) | ND3 | ND | ND | ND | |
AgPath One Step RT-PCR kit | MoMLV | 1009025 | 2/16 (13, 1–10) | ND | ND | ND | ND | |
AgPath One Step RT-PCR kit | MoMLV | 1003027 | 0/16 | ND | ND | ND | ND | |
AgPath One Step RT-PCR kit | MoMLV | 1010031 | 2/16 (13, 1–10) | ND | ND | ND | ND | |
ABI | AmpliTaq+AgPath enzyme | MoMLV | 1009030 | 7/16 (44, 1–10) | ND | ND | ND | ND |
ABI | TaqMan 1-Step Master mix | MoMLV | 1006001 | 4/16 (25, 1–10) | 0/16 | 12/16 (75) | 0/16 | 0/16 |
ABI | TaqMan RNA to CT kit | MoMLV | 802002 | 4/16 (25, 1–10) | ND | ND | ND | ND |
Agilent | Brilliant II QRT-PCR Master mix | MoMLV | 6080963 | 6/16 (38, 1–10) | 0/16 | 15/16 (94) | 0/16 | 0/16 |
Promega | Access RT-PCR system | AMV | 316238 | 0/16 | ND | ND | ND | ND |
AccessQuick RT-PCR system | AMV | 313296 | 0/16 | 0/16 | 0/16 | 0/16 | 0/16 | |
AccessQuick RT-PCR system | AMV | 316238 | 0/16 | ND | ND | ND | ND | |
Roche | Transcriptor 1-step RT-PCR kit | AMV | 12275200 | 8/16 (50, 1–10) | 0/16 | 4/16 (25) | 0/16 | 0/16 |
Transcriptor 1-step RT-PCR kit | AMV | 12513300 | 2/16 (13, 1–10) | ND | ND | ND | ND | |
Finnzymes | RobusT I RT-PCR kit | AMV | 127 | failed | 0/16 | 2/16 (13) | 0/16 | 0/16 |
The reverse transcriptase (RT) enzymes and kits were tested by quantitative protease (pro), polymerase (pol), gag and/or nested pol RT-PCR assays and results shown are number of positive tests out of the total number of replicates. Percentages for replicate testing and copies/reaction (rxn) are in parentheses.
Mouse DNA contamination was detected using mitochondrial DNA (mtDNA) and intracisternal A particle (IAP) PCR qPCR assays.
ND, not done.