Table 1.
Efficiency of long range PCR for the distinct fragments by the use of three commercially available enzymes
| Amplicon | Fragment length in bases | GC content in % | total BAC-DNA as template (100 ng) | cell-culture derived DNA as template* | |||
|---|---|---|---|---|---|---|---|
| TaK | Pro | Qia | TaK | TaK | |||
| efficiency | efficiency | efficiency | specificity (1 × 105 copies/reaction) | sensitivity (copies/reaction) | |||
| F1.1 | 7807 | 56.5 | high | low | low | low | 300 |
| F1.2 | 13,807 | 58.9 | high | – | – | – | nd |
| F1.3 | 18,015 | 56.5 | – | – | – | nd | nd |
| F2.1 | 8278 | 57.1 | high | – | – | – | nd |
| F2.2 | 13,348 | 58.8 | high | – | – | – | nd |
| F2.3 | 17,861 | 56.4 | – | – | – | nd | nd |
| F3 | 7705 | 53.7 | high | high | low | high | 30 |
| F4 | 6671 | 48.5 | high | nd | nd | high | 30 |
*two different HCMV strains (TB40E and Merlin); high and low efficiency as determined by the strength of visible bands of the correct length; high specificity as determined by the absence of unspecific bands; TaK: LA Taq Hot Start Version Polymerase; Qia: LongRange PCR Polymerase; Pro: Promega Go Taq Long; nd: not determined; “-”: no visible band or smear